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Article(id=1210676787605017578, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2021.05.04, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1613577600000, receivedDateStr=2021-02-18, revisedDate=1618416000000, revisedDateStr=2021-04-15, acceptedDate=null, acceptedDateStr=null, onlineDate=1766577439451, onlineDateStr=2025-12-24, pubDate=1622131200000, pubDateStr=2021-05-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766577439451, onlineIssueDateStr=2025-12-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766577439451, creator=13701087609, updateTime=1766577439451, updator=13701087609, issue=Issue{id=1210676785113600955, tenantId=1146029695717560320, journalId=1189873630562394117, year='2021', volume='46', issue='5', pageStart='425', pageEnd='530', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1766577438858, creator=13701087609, updateTime=1766718730270, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1211269404306838321, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1211269404306838322, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=448, endPage=455, ext={EN=ArticleExt(id=1210676787907007478, articleId=1210676787605017578, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Transcriptomics analysis of entecavir resistant hepatitis B virus stable replication cell line, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To profile transcriptome dynamics in the entecavir resistant hepatitis B virus (ETV-r HBV) stably transfected cell line, and provide fundamental data for future studies to understand mechanisms of infection using this cell line. Methods ETV-r HBV stably transfected cell lines (HepG2.A64) and the corresponding isogenic cell lines (HepG2) were used to profile the transcriptome (n=3). We used DESeq2 software to analyze the sequencing results to identify differentially expressed genes with |log2 Fold Change|>2, padj<0.05. GO and KEGG functional enrichment analysis of differential genes were performed by clusterProfiler software, and then the transcriptional and protein levels of related genes in the DNA repair pathway were detected by RT-qPCR and Western blotting. Results A total of 613 differentially expressed genes were identified in the cell lines before and after stable ETV-r HBV transfection, of which 401 were up-regulated and 212 down-regulated (padj<0.05). GO and KEGG enrichment analysis showed that differentially expressed genes were mainly involved in the inflammatory response, PI3K/Akt signal pathway, PPAR signal pathway, NF-κB signal pathway and immune-related biological processes. Through the analysis of the relevant genes in the DNA repair pathway and the verification by RT-qPCR and Western blotting, it was found that after stable transfection of ETV-r HBV, the mRNA expression of RAD52 and XRCC2 genes in the DNA repair pathway decreased by 68.96%±7.59% and 69.58%±6.32%, respectively, and the protein expression decreased by 69.93%±3.88% and 26.47%±12.7%, respectively, and the differences were statistically significant (P<0.05). Conclusion Compared with the original background cell line HepG2, a lot of genes are differentially expressed at the transcription level in HBV stable cell line, and the DNA repair-related genes in the HepG2.A64 cell line had been decreased significantly.

, correspAuthors=Yan Liu, Hao Li, authorNote=null, correspAuthorsNote=
*Li Hao, E-mail: ;
Liu Yan, E-mail:
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目的 分析恩替卡韦耐药乙型肝炎病毒(ETV-r HBV)稳定转染前后宿主细胞转录组水平的基因差异表达，为后续开展HBV感染相关机制研究提供基础数据和参考。方法 对ETV-r HBV稳定转染细胞株(HepG2.A64)及其原始背景细胞株(HepG2)进行转录组测序(n=3)，利用DESeq2软件以|log2 Fold Change|>2、padj<0.05为差异基因筛选条件对测序结果进行差异基因筛选；利用clusterProfiler软件对差异基因进行GO功能富集分析和KEGG功能富集分析。采用RT-qPCR及Western blotting分别检测HBV稳定转染前后宿主细胞DNA修复相关基因mRNA及蛋白的表达水平。结果 转录组测序结果显示，与HepG2细胞系相比，ETV-r HBV稳定转染的HepG2.A64细胞系共有613个基因存在差异表达，其中401个上调，212个下调(padj<0.05)，GO和KEGG富集分析结果显示，差异表达基因主要参与炎症反应、PI3K/Akt信号通路、PPAR信号通路、NF-κB信号通路以及免疫相关的生物学过程。RT-qPCR及Western blotting检测结果显示，与HepG2细胞系相比，HepG2.A64细胞系中RAD52和XRCC2 mRNA表达量分别下降68.96%±7.59%、69.58%±6.32%，RAD52和XRCC2蛋白表达量分别下降69.93%±3.88%、26.47%±12.7%，差异均有统计学意义(P<0.05)。结论 与原始细胞系HepG2相比，ETV-r HBV稳定转染的HepG2.A64细胞系在转录水平出现大量差异表达基因，同时HepG2.A64细胞系中DNA修复相关基因表达量出现下调。

, correspAuthors=刘妍, 李浩, authorNote=null, correspAuthorsNote=
李浩，E-mail：;
刘妍，E-mail：
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张孝璐，硕士研究生，主要从事HBV感染对宿主DNA修复系统的影响方面的研究

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张孝璐，硕士研究生，主要从事HBV感染对宿主DNA修复系统的影响方面的研究

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张孝璐，硕士研究生，主要从事HBV感染对宿主DNA修复系统的影响方面的研究

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Hepatitis B virus infection[J]. Nat Rev Dis Primers, 2018, 4: 18035., articleTitle=Hepatitis B virus infection, refAbstract=null), Reference(id=1211282636388037394, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2014, volume=384, issue=9959, pageStart=2053, pageEnd=2063, url=null, language=null, rfNumber=[2], rfOrder=1, authorNames=Trépo C, Chan HLY, Lok A, journalName=Lancet, refType=null, unstructuredReference=Trépo C, Chan HLY, Lok A. Hepatitis B virus infection[J]. Lancet, 2014, 384(9959): 2053-2063., articleTitle=Hepatitis B virus infection, refAbstract=null), Reference(id=1211282636459340564, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=1987, volume=84, issue=4, pageStart=1005, pageEnd=1009, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=Sells MA, Chen ML, Acs G, journalName=Proc Natl Acad Sci USA, refType=null, unstructuredReference=Sells MA, Chen ML, Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA[J]. Proc Natl Acad Sci USA, 1987, 84(4): 1005-1009., articleTitle=Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA, refAbstract=null), Reference(id=1211282636530643734, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=1997, volume=41, issue=8, pageStart=1715, pageEnd=1720, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=Ladner SK, Otto MJ, Barker CS, journalName=Antimicrob Agents Chemother, refType=null, unstructuredReference=Ladner SK, Otto MJ, Barker CS, et al. Inducible expression of human hepatitis B virus (HBV) in stably transfected hepatoblastoma cells: a novel system for screening potential inhibitors of HBV replication[J]. Antimicrob Agents Chemother, 1997, 41(8): 1715-1720., articleTitle=Inducible expression of human hepatitis B virus (HBV) in stably transfected hepatoblastoma cells: a novel system for screening potential inhibitors of HBV replication, refAbstract=null), Reference(id=1211282636606141208, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2006, volume=45, issue=5, pageStart=636, pageEnd=645, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=Sun DX, Nassal M, journalName=J Hepatol, refType=null, unstructuredReference=Sun DX, Nassal M. Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus[J]. J Hepatol, 2006, 45(5): 636-645., articleTitle=Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus, refAbstract=null), Reference(id=1211282636664861466, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=36, issue=9, pageStart=1284, pageEnd=1294, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=Liu W, Song H, Chen Q, journalName=Liver Int, refType=null, unstructuredReference=Liu W, Song H, Chen Q, et al. Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of nucleos(t)ide analogues[J]. Liver Int, 2016, 36(9): 1284-1294., articleTitle=Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of nucleos(t)ide analogues, refAbstract=null), Reference(id=1211282636731970332, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2018, volume=17, issue=5, pageStart=7113, pageEnd=7121, url=null, language=null, rfNumber=[7], rfOrder=6, authorNames=Liu YT, Liu W, Zhu GY, journalName=Mol Med Rep, refType=null, unstructuredReference=Liu YT, Liu W, Zhu GY, et al. Involvement of multidrug resistance protein 4 in the hepatocyte efflux of lamivudine and entecavir[J]. Mol Med Rep, 2018, 17(5): 7113-7121., articleTitle=Involvement of multidrug resistance protein 4 in the hepatocyte efflux of lamivudine and entecavir, refAbstract=null), Reference(id=1211282636799079198, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2018, volume=155, issue=null, pageStart=39, pageEnd=47, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=Liu Y, Yao W, Si L, journalName=Antiviral Res, refType=null, unstructuredReference=Liu Y, Yao W, Si L, et al. Chinese herbal extract Su-duxing had potent inhibitory effects on both wild-type and entecavir-resistant hepatitis B virus (HBV) in vitro and effectively suppressed HBV replication in mouse model[J]. Antiviral Res, 2018, 155: 39-47., articleTitle=Chinese herbal extract Su-duxing had potent inhibitory effects on both wild-type and entecavir-resistant hepatitis B virus (HBV) in vitro and effectively suppressed HBV replication in mouse model, refAbstract=null), Reference(id=1211282636874576672, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=12, issue=9, pageStart=1104, pageEnd=1113, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=Li H, Sheng C, Liu H, journalName=Int J Biol Sci, refType=null, unstructuredReference=Li H, Sheng C, Liu H, et al. An effective molecular target site in hepatitis B virus S gene for Cas9 cleavage and mutational inactivation[J]. Int J Biol Sci, 2016, 12(9): 1104-1113., articleTitle=An effective molecular target site in hepatitis B virus S gene for Cas9 cleavage and mutational inactivation, refAbstract=null), Reference(id=1211282636950074146, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2017, volume=7, issue=null, pageStart=91, pageEnd=null, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=Li H, Sheng C, Wang S, journalName=Front Cell Infect Microbiol, refType=null, unstructuredReference=Li H, Sheng C, Wang S, et al. Removal of integrated hepatitis B virus DNA using CRISPR-Cas9[J]. Front Cell Infect Microbiol, 2017, 7: 91., articleTitle=Removal of integrated hepatitis B virus DNA using CRISPR-Cas9, refAbstract=null), Reference(id=1211282637025571620, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=45, issue=12, pageStart=1595, pageEnd=1597, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=Ge LL, Yang XA, Piao JY, journalName=Chongqing Med, refType=null, unstructuredReference=Ge LL, Yang XA, Piao JY, et al. Screening of tumor metastasis related differentially expressed genes in hepatocellular carcinoma cells with FATE/BJ-HCC-2 gene stable transfection by gene chip[J]. Chongqing Med, 2016, 45(12): 1595-1597., articleTitle=Screening of tumor metastasis related differentially expressed genes in hepatocellular carcinoma cells with FATE/BJ-HCC-2 gene stable transfection by gene chip, refAbstract=null), Reference(id=1211282637101069094, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=45, issue=12, pageStart=1595, pageEnd=1597, url=null, language=null, rfNumber=[11], rfOrder=11, authorNames=葛丽丽, 杨小昂, 朴军颜, journalName=重庆医学, refType=null, unstructuredReference=[葛丽丽, 杨小昂, 朴军颜, 等. 基因芯片筛选稳定转染FATE/BJ-HCC-2基因的肝癌细胞中肿瘤转移相关差异表达基因[J]. 重庆医学, 2016, 45(12): 1595-1597.], articleTitle=基因芯片筛选稳定转染FATE/BJ-HCC-2基因的肝癌细胞中肿瘤转移相关差异表达基因, refAbstract=null), Reference(id=1211282637180760871, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2018, volume=498, issue=3, pageStart=495, pageEnd=501, url=null, language=null, rfNumber=[12], rfOrder=12, authorNames=Wu Z, Wang T, Fang M, journalName=Biochem Biophys Res Commun, refType=null, unstructuredReference=Wu Z, Wang T, Fang M, et al. MFAP5 promotes tumor progression and bone metastasis by regulating ERK/MMP signaling pathways in breast cancer[J]. Biochem Biophys Res Commun, 2018, 498(3): 495-501., articleTitle=MFAP5 promotes tumor progression and bone metastasis by regulating ERK/MMP signaling pathways in breast cancer, refAbstract=null), Reference(id=1211282637285618472, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2018, volume=507, issue=1/2/3/4, pageStart=51, pageEnd=58, url=null, language=null, rfNumber=[13], rfOrder=13, authorNames=Li QM, Zhang YQ, Jiang QL, journalName=Biochem Biophys Res Commun, refType=null, unstructuredReference=Li QM, Zhang YQ, Jiang QL. MFAP5 suppression inhibits migration/invasion, regulates cell cycle and induces apoptosis via promoting ROS production in cervical cancer[J]. Biochem Biophys Res Commun, 2018, 507(1/2/3/4): 51-58., articleTitle=MFAP5 suppression inhibits migration/invasion, regulates cell cycle and induces apoptosis via promoting ROS production in cervical cancer, refAbstract=null), Reference(id=1211282637361115945, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2015, volume=23, issue=7, pageStart=1371, pageEnd=1378, url=null, language=null, rfNumber=[14], rfOrder=14, authorNames=Vaittinen M, Kolehmainen M, Rydén M, journalName=Obesity (Silver Spring), refType=null, unstructuredReference=Vaittinen M, Kolehmainen M, Rydén M, et al. MFAP5 is related to obesity-associated adipose tissue and extracellular matrix remodeling and inflammation[J]. Obesity (Silver Spring), 2015, 23(7): 1371-1378., articleTitle=MFAP5 is related to obesity-associated adipose tissue and extracellular matrix remodeling and inflammation, refAbstract=null), Reference(id=1211282637415641898, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=40, issue=null, pageStart=382, pageEnd=393, url=null, language=null, rfNumber=[15], rfOrder=15, authorNames=Cheng Y, Wang KY, Geng LL, journalName=EBioMedicine, refType=null, unstructuredReference=Cheng Y, Wang KY, Geng LL, et al. Identification of candidate diagnostic and prognostic biomarkers for pancreatic carcinoma[J]. EBioMedicine, 2019, 40: 382-393., articleTitle=Identification of candidate diagnostic and prognostic biomarkers for pancreatic carcinoma, refAbstract=null), Reference(id=1211282637507916587, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=6, issue=null, pageStart=26605, pageEnd=null, url=null, language=null, rfNumber=[16], rfOrder=16, authorNames=Yang SF, Yeh CB, Chou YE, journalName=Sci Rep, refType=null, unstructuredReference=Yang SF, Yeh CB, Chou YE, et al. Serpin peptidase inhibitor(SERPINB5) haplotypes are associated with susceptibility to hepatocellular carcinoma[J]. Sci Rep, 2016, 6: 26605., articleTitle=Serpin peptidase inhibitor(SERPINB5) haplotypes are associated with susceptibility to hepatocellular carcinoma, refAbstract=null), Reference(id=1211282637566636844, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=16, issue=11, pageStart=2846, pageEnd=2854, url=null, language=null, rfNumber=[17], rfOrder=17, authorNames=Murphy CM, Xu YP, Li F, journalName=Cell Rep, refType=null, unstructuredReference=Murphy CM, Xu YP, Li F, et al. Hepatitis B virus X protein promotes degradation of SMC5/6 to enhance HBV replication[J]. Cell Rep, 2016, 16(11): 2846-2854., articleTitle=Hepatitis B virus X protein promotes degradation of SMC5/6 to enhance HBV replication, refAbstract=null), Reference(id=1211282637637940013, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=531, issue=7594, pageStart=386, pageEnd=389, url=null, language=null, rfNumber=[18], rfOrder=18, authorNames=Decorsière A, Mueller H, van Breugel PC, journalName=Nature, refType=null, unstructuredReference=Decorsière A, Mueller H, van Breugel PC, et al. Hepatitis B virus X protein identifies the Smc5/6 complex as a host restriction factor[J]. Nature, 2016, 531(7594): 386-389., articleTitle=Hepatitis B virus X protein identifies the Smc5/6 complex as a host restriction factor, refAbstract=null), Reference(id=1211282637700854574, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=17, issue=7, pageStart=399, pageEnd=412, url=null, language=null, rfNumber=[19], rfOrder=19, authorNames=Uhlmann F, journalName=Nat Rev Mol Cell Biol, refType=null, unstructuredReference=Uhlmann F. SMC complexes: from DNA to chromosomes[J]. Nat Rev Mol Cell Biol, 2016, 17(7): 399-412., articleTitle=SMC complexes: from DNA to chromosomes, refAbstract=null), Reference(id=1211282637763769135, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2018, volume=371, issue=2, pageStart=330, pageEnd=341, url=null, language=null, rfNumber=[20], rfOrder=20, authorNames=Zhang Z, Huo H, Liao K, journalName=Exp Cell Res, refType=null, unstructuredReference=Zhang Z, Huo H, Liao K, et al. RPA1 downregulation enhances nasopharyngeal cancer radiosensitivity via blocking RAD51 to the DNA damage site[J]. Exp Cell Res, 2018, 371(2): 330-341., articleTitle=RPA1 downregulation enhances nasopharyngeal cancer radiosensitivity via blocking RAD51 to the DNA damage site, refAbstract=null), Reference(id=1211282638942368560, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=7, issue=2, pageStart=297, pageEnd=312, url=null, language=null, rfNumber=[21], rfOrder=21, authorNames=Sekiba K, Otsuka M, Ohno M, journalName=Cell Mol Gastroenterol Hepatol, refType=null, unstructuredReference=Sekiba K, Otsuka M, Ohno M, et al. Inhibition of HBV transcription from cccDNA With nitazoxanide by targeting the HBx-DDB1 interaction[J]. Cell Mol Gastroenterol Hepatol, 2019, 7(2): 297-312., articleTitle=Inhibition of HBV transcription from cccDNA With nitazoxanide by targeting the HBx-DDB1 interaction, refAbstract=null), Reference(id=1211282639026254641, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=32, issue=10, pageStart=808, pageEnd=812, url=null, language=null, rfNumber=[22], rfOrder=22, authorNames=Gerossier L, Decorsière A, Abdul F, journalName=Med Sci(Paris), refType=null, unstructuredReference=Gerossier L, Decorsière A, Abdul F, et al. A new restriction factor against HBV infection: the SMC5/6 complex[J]. Med Sci(Paris), 2016, 32(10): 808-812., articleTitle=A new restriction factor against HBV infection: the SMC5/6 complex, refAbstract=null), Reference(id=1211282639093363506, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=172, issue=null, pageStart=104618, pageEnd=null, url=null, language=null, rfNumber=[23], rfOrder=23, authorNames=Rivière L, Quioc-Salomon B, Fallot G, journalName=Antiviral Res, refType=null, unstructuredReference=Rivière L, Quioc-Salomon B, Fallot G, et al. Hepatitis B virus replicating in hepatocellular carcinoma encodes HBx variants with preserved ability to antagonize restriction by Smc5/6[J]. Antiviral Res, 2019, 172: 104618., articleTitle=Hepatitis B virus replicating in hepatocellular carcinoma encodes HBx variants with preserved ability to antagonize restriction by Smc5/6, refAbstract=null), Reference(id=1211282639164666675, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2017, volume=36, issue=8, pageStart=693, pageEnd=708, url=null, language=null, rfNumber=[24], rfOrder=24, authorNames=Krupa R, Czarny P, Wigner P, journalName=DNA Cell Biol, refType=null, unstructuredReference=Krupa R, Czarny P, Wigner P, et al. The relationship between single-nucleotide polymorphisms, the expression of DNA damage response genes, and hepatocellular carcinoma in a Polish population[J]. DNA Cell Biol, 2017, 36(8): 693-708., articleTitle=The relationship between single-nucleotide polymorphisms, the expression of DNA damage response genes, and hepatocellular carcinoma in a Polish population, refAbstract=null), Reference(id=1211282639244358452, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2001, volume=276, issue=17, pageStart=14124, pageEnd=14132, url=null, language=null, rfNumber=[25], rfOrder=25, authorNames=Jaitovich-Groisman I, Benlimame N, Slagle BL, journalName=J Biol Chem, refType=null, unstructuredReference=Jaitovich-Groisman I, Benlimame N, Slagle BL, et al. Transcriptional regulation of the TFIIH transcription repair components XPB and XPD by the hepatitis B virus x protein in liver cells and transgenic liver tissue[J]. J Biol Chem, 2001, 276(17): 14124-14132., articleTitle=Transcriptional regulation of the TFIIH transcription repair components XPB and XPD by the hepatitis B virus x protein in liver cells and transgenic liver tissue, refAbstract=null), Reference(id=1211282639319855925, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=64, issue=1 Suppl, pageStart=S84, pageEnd=S101, url=null, language=null, rfNumber=[26], rfOrder=26, authorNames=Levrero M, Zucman-Rossi J, journalName=J Hepatol, refType=null, unstructuredReference=Levrero M, Zucman-Rossi J. Mechanisms of HBV-induced hepatocellular carcinoma[J]. J Hepatol, 2016, 64(1 Suppl): S84-S101., articleTitle=Mechanisms of HBV-induced hepatocellular carcinoma, refAbstract=null), Reference(id=1211282639399547702, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2011, volume=11, issue=null, pageStart=48, pageEnd=null, url=null, language=null, rfNumber=[27], rfOrder=27, authorNames=Qadri I, Fatima K, AbdeL-Hafiz H, journalName=BMC Microbiol, refType=null, unstructuredReference=Qadri I, Fatima K, AbdeL-Hafiz H. Hepatitis B virus X protein impedes the DNA repair via its association with transcription factor, TFIIH[J]. BMC Microbiol, 2011, 11: 48., articleTitle=Hepatitis B virus X protein impedes the DNA repair via its association with transcription factor, TFIIH, refAbstract=null), Reference(id=1211282639466656567, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=10, issue=1, pageStart=2885, pageEnd=null, url=null, language=null, rfNumber=[28], rfOrder=28, authorNames=Kokic G, Chernev A, Tegunov D, journalName=Nat Commun, refType=null, unstructuredReference=Kokic G, Chernev A, Tegunov D, et al. Structural basis of TFIIH activation for nucleotide excision repair[J]. Nat Commun, 2019, 10(1): 2885., articleTitle=Structural basis of TFIIH activation for nucleotide excision repair, refAbstract=null), Reference(id=1211282639525376824, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2017, volume=67, issue=1, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[29], rfOrder=29, authorNames=Mazina OM, Keskin H, Hanamshet K, journalName=Mol Cell, refType=null, unstructuredReference=Mazina OM, Keskin H, Hanamshet K, et al. Rad52 inverse strand exchange drives RNA-templated DNA double-strand break repair[J]. Mol Cell, 2017, 67(1): 19-29.e3., articleTitle=Rad52 inverse strand exchange drives RNA-templated DNA double-strand break repair, refAbstract=null), Reference(id=1211282639592485689, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=11, issue=11, pageStart=1622, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=30, authorNames=Nogueira A, Fernandes M, Catarino R, journalName=Cancers (Basel), refType=null, unstructuredReference=Nogueira A, Fernandes M, Catarino R, et al. RAD52 functions in homologous recombination and its importance on genomic integrity maintenance and cancer therapy[J]. Cancers (Basel), 2019, 11(11): 1622., articleTitle=RAD52 functions in homologous recombination and its importance on genomic integrity maintenance and cancer therapy, refAbstract=null), Reference(id=1211282639667983162, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=38, issue=7, pageStart=504, pageEnd=509, url=null, language=null, rfNumber=[31], rfOrder=31, authorNames=Li HM, Yuan P, Yu DK, journalName=Chin J Oncol, refType=null, unstructuredReference=Li HM, Yuan P, Yu DK, et al. Genetic variation in DNA repair gene RAD52 is associated with the response to platinum-based chemotherapy in SCLC patients[J]. Chin J Oncol, 2016, 38(7): 504-509., articleTitle=Genetic variation in DNA repair gene RAD52 is associated with the response to platinum-based chemotherapy in SCLC patients, refAbstract=null), Reference(id=1211282639730897723, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=64, issue=6, pageStart=1127, pageEnd=1134, url=null, language=null, rfNumber=[32], rfOrder=32, authorNames=Sotiriou SK, Kamileri I, Lugli N, journalName=Mol Cell, refType=null, unstructuredReference=Sotiriou SK, Kamileri I, Lugli N, et al. Mammalian RAD52 functions in break-induced replication repair of collapsed DNA replication forks[J]. Mol Cell, 2016, 64(6): 1127-1134., articleTitle=Mammalian RAD52 functions in break-induced replication repair of collapsed DNA replication forks, refAbstract=null), Reference(id=1211282639789617980, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2019, volume=23, issue=1, pageStart=1, pageEnd=7, url=null, language=null, rfNumber=[33], rfOrder=33, authorNames=Andreassen PR, Hanenberg H, journalName=Atlas Genet Cytogenet Oncol Haematol, refType=null, unstructuredReference=Andreassen PR, Hanenberg H. XRCC2 (X-ray repair cross complementing 2)[J]. Atlas Genet Cytogenet Oncol Haematol, 2019, 23(1): 1-7., articleTitle=XRCC2 (X-ray repair cross complementing 2), refAbstract=null), Reference(id=1211282639852532541, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=53, issue=10, pageStart=672, pageEnd=680, url=null, language=null, rfNumber=[34], rfOrder=34, authorNames=Park JY, Virts EL, Jankowska A, journalName=J Med Genet, refType=null, unstructuredReference=Park JY, Virts EL, Jankowska A, et al. Complementation of hypersensitivity to DNA interstrand crosslinking agents demonstrates that XRCC2 is a Fanconi anaemia gene[J]. J Med Genet, 2016, 53(10): 672-680., articleTitle=Complementation of hypersensitivity to DNA interstrand crosslinking agents demonstrates that XRCC2 is a Fanconi anaemia gene, refAbstract=null), Reference(id=1211282639915447102, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, doi=null, pmid=null, pmcid=null, year=2016, volume=37, issue=9, pageStart=914, pageEnd=925, url=null, language=null, rfNumber=[35], rfOrder=35, authorNames=Hilbers FS, Luijsterburg MS, Wiegant WW, journalName=Hum Mutat, refType=null, unstructuredReference=Hilbers FS, Luijsterburg MS, Wiegant WW, et al. Functional analysis of missense variants in the putative breast cancer susceptibility gene XRCC2[J]. Hum Mutat, 2016, 37(9): 914-925., articleTitle=Functional analysis of missense variants in the putative breast cancer susceptibility gene XRCC2, refAbstract=null)], funds=[Fund(id=1211282635989578503, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, awardId=81573676, language=EN, fundingSource=National Natural Science Foundation of China(81573676), fundOrder=null, country=null), Fund(id=1211282636056687369, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, awardId=81573676, language=CN, fundingSource=国家自然科学基金(81573676), fundOrder=null, country=null), Fund(id=1211282636123796235, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, awardId=7194302, language=EN, fundingSource=Natural Science Foundation of Beijing(7194302), fundOrder=null, country=null), Fund(id=1211282636182516493, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, awardId=7194302, language=CN, fundingSource=北京市自然科学基金(7194302), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1211282630922859152, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, xref=1, ext=[AuthorCompanyExt(id=1211282630931247761, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, companyId=1211282630922859152, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1State Key Laboratory of Pathogenic Microorganism Biosafety, Institute of Microbiology and Epidemiology, Academy of Military Medicine, Academy of Military Sciences, Beijing 100071, China), AuthorCompanyExt(id=1211282630943830674, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, companyId=1211282630922859152, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1病原微生物生物安全国家重点实验室，军事科学院军事医学研究院微生物流行病研究所，北京 100071)]), AuthorCompany(id=1211282630998356629, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, xref=2, ext=[AuthorCompanyExt(id=1211282631002550933, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, companyId=1211282630998356629, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2Department of Infectious Diseases, the Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China), AuthorCompanyExt(id=1211282631015133847, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, companyId=1211282630998356629, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2解放军总医院第五医学中心感染病医学部，北京 100039)])], figs=[ArticleFig(id=1211282635012305646, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Fig.1, caption=Detection of hepatitis B markers in HepG2 and HepG2.A64 cells, figureFileSmall=g6LN5CWBTMQQ1me+l0FohQ==, figureFileBig=HpkKbn8seuC2f5D+lshuYQ==, tableContent=null), ArticleFig(id=1211282635087803120, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=图1, caption=HepG2、HepG2.A64细胞的乙肝标志物检测

S表示样本的OD值，CO值表示检测临界值；与HepG2细胞系比较，(1)P<0.05。

, figureFileSmall=g6LN5CWBTMQQ1me+l0FohQ==, figureFileBig=HpkKbn8seuC2f5D+lshuYQ==, tableContent=null), ArticleFig(id=1211282635171689202, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Fig.2, caption=Volcano map and cluster heat map of differentially expressed genes, figureFileSmall=SbN1pS7u/JG7P3YtRw8i3w==, figureFileBig=3S5IRkzOoz0dm/oY9wZYDA==, tableContent=null), ArticleFig(id=1211282635238798068, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=图2, caption=差异表达基因的火山图及聚类热图

A. 两细胞系差异表达基因的火山图，红点表示表达量上调的基因，绿点表示表达量下调的基因，蓝点表示表达量差异无统计学意义的基因；B. 两细胞系差异表达基因的聚类热图，蓝色表示基因表达量下调，红色表示基因表达量上调

, figureFileSmall=SbN1pS7u/JG7P3YtRw8i3w==, figureFileBig=3S5IRkzOoz0dm/oY9wZYDA==, tableContent=null), ArticleFig(id=1211282635318489846, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Fig.3, caption=Enrichment analysis of differentially expressed genes, figureFileSmall=Y6daHqum4Zq2I90LJhItoA==, figureFileBig=lr253u+a/hAq0QP3g3IELg==, tableContent=null), ArticleFig(id=1211282635377210104, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=图3, caption=差异表达基因富集分析

A. GO功能富集分析中差异显著基因的表达量，横坐标表示差异显著的基因，纵坐标表示转录水平基因表达值(即FPKM值)；B. 差异表达基因KEGG功能富集分析，横坐标表示注释到KEGG通路上的差异基因数，纵坐标表示KEGG通路名称

, figureFileSmall=Y6daHqum4Zq2I90LJhItoA==, figureFileBig=lr253u+a/hAq0QP3g3IELg==, tableContent=null), ArticleFig(id=1211282635448513274, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Fig.4, caption=RT-qPCR and Western blotting analysis of related genes of HepG2.A64 and HepG2 cells in DNA repair pathway, figureFileSmall=tiSUS46qST+JW8qgoW7SuA==, figureFileBig=dP2jnumoOgjKTjeiyfLiMQ==, tableContent=null), ArticleFig(id=1211282635511427836, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=图4, caption=两细胞系DNA修复通路中相关基因的RT-qPCR和Western blotting检测结果

A. mRNA相对表达量；B. Western blotting检测(红框示表达下调基因)；C. 蛋白相对表达量；与HepG2细胞系比较，(1)P<0.05。

, figureFileSmall=tiSUS46qST+JW8qgoW7SuA==, figureFileBig=dP2jnumoOgjKTjeiyfLiMQ==, tableContent=null), ArticleFig(id=1211282635578536702, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Tab.1, caption=

Primers sequence for RT-qPCR

, figureFileSmall=null, figureFileBig=null, tableContent=
基因引物序列
RAD51上游：5'-ATTGACCGGGGTGGAGGTGA-3'
下游：5'-AGCTGGGTCTGGTGGTCTGT-3'
RAD52上游：5'-AGCCATGCTGTGATACCGGC-3'
下游：5'-GGCGTGGAGACTCGAACCTG-3'
XRCC2上游：5'-ATGGGCAAAAGAGACCGAG-3'
下游：5'-CTGACTCATCCACCTGCTTC-3'
CETN2上游：5'-GCATCAAGTTCTCAGCGAAAAAG-3'
下游：5'-CCATCCGCATCGAAAAGATCA-3'
DDB1上游：5'-GATCATCCGGAATGGAATTG-3'
下游：5'-ATTAGGGTCAGACCGCAGTG-3'
RPA1上游：5'-AAGTGGAGACCTACAACGAC-3'
下游：5'-ACAACCACCTGAGCGTAT-3'
MCM4上游：5'-TGAACCTCTATACATGCAACGAC-3'
下游：5'-CAGGGTAACGGTCAAAGAAGATT-3'
MCM9上游：5'-TGAGCAGTATTACACCTTTTGCC-3'
下游：5'-GTTGGAGACGAAGACAAGCCT-3'
MLH1上游：5'-GCAAACCCCTGTCCAGTCAG-3'
下游：5'-CTGGGAGTTCAAGCATCTCCT-3'
SMC5上游：5'-CGTGGAAGGCTCTATCGTCC-3'
下游：5'-GCTCGACTTCCCTGTTCCATT-3'
SMC6上游：5'-TGGCAACAATGGAAGTGGGA-3'
下游：5'-CAAGACCGACTATGAGAGCTGTGA-3'
β-actin上游：5'-CCTGGCACCCAGCACAA-3'
下游：5'-GCCGATCCACACGGAGTACT-3'
), ArticleFig(id=1211282635649839872, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=表1, caption=

RT-qPCR引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
基因引物序列
RAD51上游：5'-ATTGACCGGGGTGGAGGTGA-3'
下游：5'-AGCTGGGTCTGGTGGTCTGT-3'
RAD52上游：5'-AGCCATGCTGTGATACCGGC-3'
下游：5'-GGCGTGGAGACTCGAACCTG-3'
XRCC2上游：5'-ATGGGCAAAAGAGACCGAG-3'
下游：5'-CTGACTCATCCACCTGCTTC-3'
CETN2上游：5'-GCATCAAGTTCTCAGCGAAAAAG-3'
下游：5'-CCATCCGCATCGAAAAGATCA-3'
DDB1上游：5'-GATCATCCGGAATGGAATTG-3'
下游：5'-ATTAGGGTCAGACCGCAGTG-3'
RPA1上游：5'-AAGTGGAGACCTACAACGAC-3'
下游：5'-ACAACCACCTGAGCGTAT-3'
MCM4上游：5'-TGAACCTCTATACATGCAACGAC-3'
下游：5'-CAGGGTAACGGTCAAAGAAGATT-3'
MCM9上游：5'-TGAGCAGTATTACACCTTTTGCC-3'
下游：5'-GTTGGAGACGAAGACAAGCCT-3'
MLH1上游：5'-GCAAACCCCTGTCCAGTCAG-3'
下游：5'-CTGGGAGTTCAAGCATCTCCT-3'
SMC5上游：5'-CGTGGAAGGCTCTATCGTCC-3'
下游：5'-GCTCGACTTCCCTGTTCCATT-3'
SMC6上游：5'-TGGCAACAATGGAAGTGGGA-3'
下游：5'-CAAGACCGACTATGAGAGCTGTGA-3'
β-actin上游：5'-CCTGGCACCCAGCACAA-3'
下游：5'-GCCGATCCACACGGAGTACT-3'
), ArticleFig(id=1211282635733725954, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=EN, label=Tab.2, caption=

Characteristics of differentially expressed genes in HepG2.A64 cells compared to HepG2 cells

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称FPKMlog2(差异倍数)padj调节类型
HepG2细胞HepG2.A64细胞
CGA1.778104.7175.893 384<0.001上调
A2M0.40489.7717.794 234<0.001上调
MFAP522.3890.111–7.640 850<0.001下调
IGFBP420.9710.472–5.471 180<0.001下调
SERPINB515.5920.353–5.451 520<0.001下调
ALPI586.95225.409–4.524 290<0.001下调
MAGEA36.05860.1783.323 125<0.001上调
COL15A145.4025.651–2.992 120<0.001下调
CTGF1.55219.2143.637 548<0.001上调
COL4A29.4191.488–2.651 430<0.001下调
GPC51.18910.6553.170 432<0.001上调
ALPP1644.843195.210–3.060 210<0.001下调
), ArticleFig(id=1211282635830194948, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676787605017578, language=CN, label=表2, caption=

HepG2.A64细胞与HepG2细胞差异表达基因的特征

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称FPKMlog2(差异倍数)padj调节类型
HepG2细胞HepG2.A64细胞
CGA1.778104.7175.893 384<0.001上调
A2M0.40489.7717.794 234<0.001上调
MFAP522.3890.111–7.640 850<0.001下调
IGFBP420.9710.472–5.471 180<0.001下调
SERPINB515.5920.353–5.451 520<0.001下调
ALPI586.95225.409–4.524 290<0.001下调
MAGEA36.05860.1783.323 125<0.001上调
COL15A145.4025.651–2.992 120<0.001下调
CTGF1.55219.2143.637 548<0.001上调
COL4A29.4191.488–2.651 430<0.001下调
GPC51.18910.6553.170 432<0.001上调
ALPP1644.843195.210–3.060 210<0.001下调
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恩替卡韦耐药乙型肝炎病毒稳定复制细胞系的转录组学分析
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张孝璐 1 , 杨兰 1 , 孙岩松 1 , 徐东平 2 , 李晓峰 1 , 刘妍 2, * , 李浩 1, *
解放军医学杂志 | 基础研究 2021,46(5): 448-455
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解放军医学杂志 | 基础研究 2021, 46(5): 448-455
恩替卡韦耐药乙型肝炎病毒稳定复制细胞系的转录组学分析
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张孝璐1, 杨兰1, 孙岩松1, 徐东平2, 李晓峰1, 刘妍2, * , 李浩1, *
作者信息
  • 1病原微生物生物安全国家重点实验室，军事科学院军事医学研究院微生物流行病研究所，北京 100071
  • 2解放军总医院第五医学中心感染病医学部，北京 100039

    • 张孝璐，硕士研究生，主要从事HBV感染对宿主DNA修复系统的影响方面的研究

通讯作者:

李浩，E-mail：;
刘妍，E-mail：
Transcriptomics analysis of entecavir resistant hepatitis B virus stable replication cell line
Xiao-Lu Zhang1, Lan Yang1, Yan-Song Sun1, Dong-Ping Xu2, Xiao-Feng Li1, Yan Liu2, * , Hao Li1, *
Affiliations
  • 1State Key Laboratory of Pathogenic Microorganism Biosafety, Institute of Microbiology and Epidemiology, Academy of Military Medicine, Academy of Military Sciences, Beijing 100071, China
  • 2Department of Infectious Diseases, the Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China
出版时间: 2021-05-28 doi: 10.11855/j.issn.0577-7402.2021.05.04
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摘要
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目的 分析恩替卡韦耐药乙型肝炎病毒(ETV-r HBV)稳定转染前后宿主细胞转录组水平的基因差异表达，为后续开展HBV感染相关机制研究提供基础数据和参考。方法 对ETV-r HBV稳定转染细胞株(HepG2.A64)及其原始背景细胞株(HepG2)进行转录组测序(n=3)，利用DESeq2软件以|log2 Fold Change|>2、padj<0.05为差异基因筛选条件对测序结果进行差异基因筛选；利用clusterProfiler软件对差异基因进行GO功能富集分析和KEGG功能富集分析。采用RT-qPCR及Western blotting分别检测HBV稳定转染前后宿主细胞DNA修复相关基因mRNA及蛋白的表达水平。结果 转录组测序结果显示，与HepG2细胞系相比，ETV-r HBV稳定转染的HepG2.A64细胞系共有613个基因存在差异表达，其中401个上调，212个下调(padj<0.05)，GO和KEGG富集分析结果显示，差异表达基因主要参与炎症反应、PI3K/Akt信号通路、PPAR信号通路、NF-κB信号通路以及免疫相关的生物学过程。RT-qPCR及Western blotting检测结果显示，与HepG2细胞系相比，HepG2.A64细胞系中RAD52和XRCC2 mRNA表达量分别下降68.96%±7.59%、69.58%±6.32%，RAD52和XRCC2蛋白表达量分别下降69.93%±3.88%、26.47%±12.7%，差异均有统计学意义(P<0.05)。结论 与原始细胞系HepG2相比，ETV-r HBV稳定转染的HepG2.A64细胞系在转录水平出现大量差异表达基因，同时HepG2.A64细胞系中DNA修复相关基因表达量出现下调。

乙型肝炎病毒  /  DNA修复  /  计算生物学  /  转录组测序  /  恩替卡韦  /  HepG2.A64细胞系

Objective To profile transcriptome dynamics in the entecavir resistant hepatitis B virus (ETV-r HBV) stably transfected cell line, and provide fundamental data for future studies to understand mechanisms of infection using this cell line. Methods ETV-r HBV stably transfected cell lines (HepG2.A64) and the corresponding isogenic cell lines (HepG2) were used to profile the transcriptome (n=3). We used DESeq2 software to analyze the sequencing results to identify differentially expressed genes with |log2 Fold Change|>2, padj<0.05. GO and KEGG functional enrichment analysis of differential genes were performed by clusterProfiler software, and then the transcriptional and protein levels of related genes in the DNA repair pathway were detected by RT-qPCR and Western blotting. Results A total of 613 differentially expressed genes were identified in the cell lines before and after stable ETV-r HBV transfection, of which 401 were up-regulated and 212 down-regulated (padj<0.05). GO and KEGG enrichment analysis showed that differentially expressed genes were mainly involved in the inflammatory response, PI3K/Akt signal pathway, PPAR signal pathway, NF-κB signal pathway and immune-related biological processes. Through the analysis of the relevant genes in the DNA repair pathway and the verification by RT-qPCR and Western blotting, it was found that after stable transfection of ETV-r HBV, the mRNA expression of RAD52 and XRCC2 genes in the DNA repair pathway decreased by 68.96%±7.59% and 69.58%±6.32%, respectively, and the protein expression decreased by 69.93%±3.88% and 26.47%±12.7%, respectively, and the differences were statistically significant (P<0.05). Conclusion Compared with the original background cell line HepG2, a lot of genes are differentially expressed at the transcription level in HBV stable cell line, and the DNA repair-related genes in the HepG2.A64 cell line had been decreased significantly.

hepatitis B virus  /  DNA repair  /  computational biology  /  transcriptome sequencing  /  entecavir  /  HepG2.A64 cell line
张孝璐, 杨兰, 孙岩松, 徐东平, 李晓峰, 刘妍, 李浩. 恩替卡韦耐药乙型肝炎病毒稳定复制细胞系的转录组学分析. 解放军医学杂志, 2021 , 46 (5) : 448 -455 . DOI: 10.11855/j.issn.0577-7402.2021.05.04
Xiao-Lu Zhang, Lan Yang, Yan-Song Sun, Dong-Ping Xu, Xiao-Feng Li, Yan Liu, Hao Li. Transcriptomics analysis of entecavir resistant hepatitis B virus stable replication cell line[J]. Medical Journal of Chinese People’s Liberation Army, 2021 , 46 (5) : 448 -455 . DOI: 10.11855/j.issn.0577-7402.2021.05.04
正文
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乙型肝炎病毒(hepatitis B virus，HBV)是引起急性及慢性乙型肝炎的重要病原体[1]。全世界约有2.91亿人感染HBV，每年约有100万人死于HBV引发的肝硬化或肝细胞癌(hepatocellular carcinoma，HCC)，占全球肝癌患者总数的50%以上[2]。为研究HBV的感染机制及生物学特性，近年来研究者们已建立了多种稳定表达HBV病毒颗粒的细胞系，如HepG2.2.15细胞系[3]、HepAD38细胞系[4]、Huh7.93细胞系等[5]。经典的HepG2.2.15细胞系已经建立30余年，其宿主基因与原始HepG2细胞有很大差别，而这种差别可能是由于宿主基因组突变长时间累积导致的。2011年，Liu等[6]基于我国流行的C基因型恩替卡韦(entecavir，ETV)耐药HBV病毒株，建立了HBV稳定复制细胞系HepG2.A64(CCTCC C 201163)，维持在含380 μg/ml遗传霉素(G418)的高糖DMEM培养基中，可持续44个月以上产生乙型肝炎病毒表面抗原(HBsAg)及HBV DNA。由于该细胞系可产生更多的病毒标志物及HBV cccDNA，且更易于培养及转染，已成为研究耐药HBV感染机制、生物学特性、药物疗效、治疗方案、新药及疫苗的又一个相对理想的模型。然而目前针对HBV稳定转染的HepG2.A64细胞系转录组层面的研究尚少见。本研究利用高通量转录组测序技术，对HepG2细胞与恩替卡韦耐药乙型肝炎病毒(ETV-r HBV)稳定转染的HepG2.A64细胞转录组测序数据进行对比分析，探讨HBV稳定转染对宿主细胞DNA修复相关基因表达的影响。
1 材料与方法
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1.1 主要材料、试剂及仪器
人肝癌细胞系HepG2细胞(ATCC® HB-8065)购自美国ATCC公司；ETV-r HBV稳定转染的HepG2.A64细胞系由解放军总医院第五医学中心自主研发构建(CCTCC C 201163)，国家发明专利号ZL201010586554.6。EallBio胎牛血清购自北京中生奥邦生物科技有限公司；胰酶、青霉素、链霉素、L-谷氨酰胺均购自美国Gibco公司；HBsAg酶联免疫吸附(ELISA)检测试剂盒购自北京万泰生物药业股份有限公司；HBV核酸测定试剂盒(PCR-荧光探针法)购自中山大学达安基因股份有限公司；细胞RNA提取试剂盒(74136)购自德国QIAGEN公司；“一步法”实时荧光定量PCR(RT-qPCR)试剂盒(RR066A)购自日本TaKaRa公司；BCA蛋白测定试剂盒(A53226)购自美国赛默飞世尔科技公司；抗DNA损伤结合蛋白1(DDB1)抗体(ab133534)、抗X线修复交叉互补基因2(XRCC2)抗体(ab124900)、抗复制蛋白A1(RPA1)抗体(ab79398)、抗辐射敏感51(RAD51)抗体(ab133534)、抗辐射敏感52(RAD52)抗体(ab124971)、抗β-肌动蛋白(β-actin)抗体(ab6276)、山羊抗小鼠IgG抗体(ab205719)购自英国Abcam公司，山羊抗兔IgG抗体(GTX213110-01)购自美国GeneTex公司；荧光定量PCR仪(Mastercycler ep realplex)购自德国Eppendrof公司；Millipore发光底物HRP(ECL发光液，WBKLS0500)购自美国Millipore公司；蛋白电泳仪(PowerPacTM基础电泳仪)购自美国Bio-Rad公司；Amersham Imager 600凝胶成像系统购自美国GE公司。
1.2 实验方法
1.2.1 细胞培养
HepG2、HepG2.A64细胞均培养于含1%青霉素和链霉素、1% L-谷氨酰胺及10%胎牛血清的高糖DMEM培养基中，置于37℃、含5%CO2的细胞培养箱中培养。待细胞融合至90%时弃掉细胞上清，用2 ml PBS缓冲液洗涤2次后加1 ml胰酶消化，待显微镜下观察到细胞变圆、不再连接成片时，加2 ml培养基终止消化。轻轻吹打培养瓶底部至所有细胞脱落，1000 r/min离心5 min，弃上清，加3 ml培养基重悬混匀细胞，取1 ml细胞悬液至培养瓶并补加4 ml培养基吹打混匀，置于37℃、含5% CO2的细胞培养箱中培养。
1.2.2 HBV标志物检测
取对数生长期HepG2、HepG2.A64细胞，调整细胞浓度至1×105个/ml，将1 ml细胞传代至25 cm2细胞培养瓶中，48 h后收集细胞培养基，10 000 r/min离心5 min，收集细胞培养基上清2 ml。随后以相同细胞量进行细胞传代，连续培养4代。取100 μl细胞上清液，采用ELISA法检测HBsAg的含量，按照试剂盒说明书进行操作，ELISA法检测设定：cut-off(CO)值=阴性对照样本的OD值×2.1(OD值表示样本在450 nm波长下的光密度值，阴性对照样本的OD值低于0.05者按0.05计算)，样品S/CO值≥1.0为阳性(S表示样本的OD值，CO值表示检测临界值)；HBV DNA定量检测按照HBV核酸测定试剂盒(PCR-荧光探针法)说明书进行，根据标准品绘制的标准曲线计算样品中HBV DNA的浓度。
1.2.3 RNA提取及测序
用Trizol法从HepG2细胞及HepG2.A64细胞中提取RNA(n=3)，用Agilent 2100生物分析仪检测RNA完整性，通过Oligo(dT)磁珠富集带有polyA尾的mRNA，随后在NEB片段化缓冲液(NEB Fragmentation Buffer)中将mRNA随机打断进行双链cDNA合成。纯化后的cDNA经过末端修复、加A、接头，并用AMPure XP beads核酸纯化试剂盒筛选250~300 bp的cDNA，经PCR扩增及产物纯化后获得文库。使用Qubit2.0荧光计对文库进行初步定量，并稀释至1.5 ng/μl，随后使用Agilent 2100生物分析仪对文库的插入片段大小(insert size)进行检测，insert size符合预期后，借助Illumina测序平台的Novaseq 6000进行RNA测序(RNA-seq)。
1.2.4 测序数据分析
采用Fastp(V0.19.4)软件对测序获得的原始数据进行过滤，使用Hisat2(2.0.5)软件将得到的高质量序列数据(clean reads)与人参考基因组(GRCH38)进行比对，并借助Subread软件中的feature Counts(1.5.0-p3)工具统计每个基因的reads数，完成基因表达的定量分析。以GRCH38为参考基因组，|log2 Fold Change|>2、padj<0.05作为差异基因筛选条件，借助DESeq2(1.16.1)标准化方法统计差异基因并绘制差异基因火山图；采用edgeR(3.18.1)软件统计每个基因上的片段数作为基因的原始表达量，并用每千个碱基的转录每百万映射读取的片段(FPKM值)表示转录水平的基因表达值；clusterProfiler(3.4.4)软件基于超几何分布原理，对所有差异基因集注释到GO或KEGG数据库的基因集并进行富集，其中padj<0.05的GO及KEGG词条为显著性富集项，padj<0.05的GO及KEGG通路为显著性富集通路。
1.2.5 RT-qPCR检测DNA修复相关基因mRNA的表达
利用primer design (https://www.ncbi.nlm.nih.gov/tools/primer-blast/)网站针对DNA修复相关通路(同源重组修复通路、非同源末端连接、核酸切除修复)中的代表性基因设计上、下游引物，序列见表1。利用QIAGEN公司的细胞RNA提取试剂盒提取HepG2、HepG2.A64细胞的RNA，将细胞RNA稀释至同一浓度(50 ng/μl)，采用“一步法”RT-qPCR试剂盒检测HepG2、HepG2.A64细胞中DNA修复相关基因的相对表达量，以β-actin作为内参照，用2–ΔΔCt法计算检测结果，用GraPhad Prism6软件作图。
1.2.6 Western blotting检测DNA修复相关基因蛋白表达水平
利用含1%蛋白酶抑制剂的RIPA裂解液裂解HepG2和HepG2.A64细胞，使用BCA蛋白测定试剂盒对裂解得到的细胞蛋白进行定量，随后进行聚丙烯酰胺凝胶电泳(SDS-PAGE)分离，并在100 mA恒定电流下将蛋白转移至甲醇激活过的PVDF膜上(35 min)。利用5%脱脂奶粉稀释一抗，各抗体稀释比例分别为DDB1(1:10 000)、XRCC2(1:10 000)、RPA1(1:1000)、RAD51(1:1000)、RAD52(1:500)、β-actin(1:5000)，一抗与PVDF膜4℃孵育过夜。取出PVDF膜，用TBST洗涤一抗，8 min/次，共4次；用5%脱脂奶粉稀释HRP标记的山羊抗兔IgG抗体(1:5000)及山羊抗小鼠IgG抗体(1:5000)，室温下与PVDF膜孵育1 h，取出PVDF膜，用TBST洗涤一抗，8 min/次，共4次；用Millipore发光底物HRP(ECL发光液)洗膜，然后在Amersham Imager 600凝胶成像系统中显影。利用Image J软件对显影条带进行灰度值统计，将目的蛋白的灰度值除以内参蛋白的灰度值以校正误差，所得结果代表目的基因蛋白相对表达量。
1.3 统计学处理
采用SPSS 25.0软件进行统计分析。计量资料以$\bar{x}±s$表示，两细胞系差异基因的比较采用t检验，P<0.05为差异有统计学意义。
2 结果
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2.1 HBV稳定转染细胞系鉴定
ELISA检测及HBV DNA荧光探针法定量检测结果显示，HepG2.A64细胞系中的HBV能够持续高水平复制并在细胞上清中稳定表达HBsAg、HBV DNA。在连续4代的培养过程中，HepG2.A64细胞上清中HBsAg含量的S/CO值维持在5.89±0.86，HBV DNA的含量维持在(12212.75±112.57) IU/ml，而HepG2细胞系的细胞培养上清中未检出HBV标志物(图1)。
2.2 HBV稳定转染后宿主细胞转录组的基因表达变化
转录组测序结果显示，与HepG2细胞系相比，ETV-r HBV稳定转染的HepG2.A64细胞系共有613个基因存在差异表达，其中401个上调，212个下调；有26 447个基因表达差异无统计学意义(图2A)。对差异基因进行聚类分析，结果显示，ETV-r HBV稳定转染的HepG2.A64细胞系表达下调的基因主要集中在MAPK信号通路、DNA复制、细胞周期相关通路中，表达上调的基因主要集中在肝细胞癌、P53信号通路、Wnt信号通路、泛素介导的蛋白水解等相关通路中(图2B)。
2.3 HBV稳定转染后宿主细胞差异表达基因的分析及功能富集
利用GO功能富集及KEGG通路富集对差异表达基因进行富集分析。GO富集分析结果显示，表达水平变化具有显著差异的前5个基因Terms主要与炎症反应、信号转导受体活性调节、代谢等生物学功能相关。将这些通路所涉及的基因按照padj值从小到大排列，选择差异表达最显著的前12个基因(图3A、表2)。其中，与HepG2细胞系相比，ETV-r HBV稳定转染的HepG2.A64细胞系中肠碱性磷酸酶(intestinal alkaline phosphatase，ALPI)、胎盘碱性磷酸酶(placental alkaline phosphatase，ALPP)、微纤维相关蛋白5(microfibril associated protein 5，MFAP5)、胰岛素样生长因子结合蛋白4(insulin-like growth factor binding protein 4，IGFBP4)、丝氨酸蛋白酶抑制剂B5(serine protease inhibitor B5，SERPINB5)、Ⅳ型胶原α2链(collagen type Ⅳ alpha 2 chain，COL4A2)、ⅩⅤ型胶原α1链(collagen type ⅩⅤ alpha 1 chai，COL15A1)等基因在转录水平的表达量明显降低，嗜铬粒蛋白A(chromogranin A，CGA)、α2-巨球蛋白基因(α2-macroglobulin，A2M)、黑色素瘤抗原基因A3(melanoma-associated antigens A3，MAGEA3)、结缔组织生长因子(connective tissue growth factor，CTGF)、磷脂酰肌醇蛋白聚糖家族成员5(glypican 5，GPC5)等基因在转录水平的表达量明显升高，差异均有统计学意义(均P<0.05)。KEGG富集分析结果显示，HBV感染后宿主细胞差异表达基因显著富集在PI3K/Akt信号通路、人乳头瘤病毒感染、细胞黏附分子、PPAR信号通路、NF-κB信号通路以及免疫相关通路中(图3B)，提示HBV感染可能影响了细胞分化、发育、代谢(糖类、脂质、蛋白质)，以及高等生物的肿瘤发生等过程。
2.4 HBV稳定转染后宿主细胞DNA修复相关基因表达的变化
RT-qPCR检测结果显示，与HepG2细胞相比，HepG2.A64细胞中RAD51 mRNA表达量下降62.25%±5.03%，RAD52 mRNA表达量下降68.96%±7.59%，XRCC2 mRNA表达量下降69.58%±6.32%，DDB1 mRNA表达量下降64.0%±9.41%，差异均有统计学意义(P<0.05，图4A)。Western blotting检测结果显示，与HepG2细胞相比，HepG2.A64细胞中RAD52、XRCC2蛋白表达下调，其中RAD52蛋白表达量下降69.93%±3.88%，XRCC2蛋白表达量下降26.47%±12.7%，差异均有统计学意义(P<0.05，图4B、4C)。
3 讨论
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HepG2.A64细胞系作为基于ETV耐药HBV病毒株建立的稳定表达HBV的细胞模型，目前已被用于抗病毒机制、病毒感染与耐药机制等方面的研究。例如，Liu等[6]利用HepG2.A64细胞系探究多药耐药蛋白4(multidrug resistance protein 4，MRP4)在核苷类似物抗反转录病毒治疗中的作用机制，证实联合应用核苷类似物和多药耐药蛋白抑制剂可减少核苷类似物的长期使用剂量；Liu等[7]利用HepG2.A64细胞系探究MPR4在拉米夫定和ETV转运过程中的作用；Liu等[8]利用HepG2.A64评价中药提取物肃毒星的抗HBV效应；本课题组前期利用HepG2.A64细胞系评估成簇的规律间隔的短回文重复序列(clustered regularly interspaced short palindromic repeats，CRISPR)对HBV的清除作用[9]，并发现CRISPR清除HBV后HepG2.A64细胞中发生了大量染色体突变恢复的现象[10]。
本研究利用高通量转录组测序技术发现，与HepG2细胞系相比，ETV-r HBV稳定转染后的HepG2.A64细胞系中共有613个基因存在差异表达，且差异表达基因主要存在于炎症反应、PPAR信号通路、NF-κB信号通路、免疫相关通路中。其中，ALPP、MFAP5、SERPINB5、ALPI、IGFBP4、COL4A2、COL15A1等基因在转录水平的表达量显著降低。而此前研究人员已在卵巢腺癌、浆液性囊腺癌和其他卵巢癌细胞中检测到ALPP基因的强异位表达，另外利用基因芯片技术筛选的稳定转染FATE/BJ-HCC-2基因的肝癌细胞中ALPP基因表达水平也明显下降[11]。MFAP5是细胞外弹性微纤维的组成部分，主要在组织发育过程中起作用[12-13]，并具有调节炎症反应的作用[14]。SERPINB5基因作为胰腺癌的生物标志物[15]，可阻止肿瘤细胞的生长、侵袭和转移[16]。本研究结果为今后HBV感染及肝癌发生中相关基因的作用机制研究提供了参考。
结合以往有关HBV感染与宿主细胞DNA修复的研究[17-20]，笔者推测本研究中ETV-r HBV稳定转染的HepG2.A64细胞系在转录水平出现大量差异表达基因可能与HBV感染抑制了宿主细胞的DNA修复能力有关。Sekiba等[21]研究证实，HBx蛋白能够与DDB1亚基相互作用，从而干扰DNA的修复能力，也可通过E3泛素连接酶靶向降解用于维持染色体结构稳定性的Smc5/6复合体，解除Smc5/6复合体对HBV基因组转录、复制的抑制作用[17-19,22-23]。同时，HBx蛋白在细胞质中可通过劫持p53基因而与细胞核中的DNA修复相关基因(如DDB1、XPB、XPD和TFIIH)发生相互作用[24]，其中HBx与TFIIH组分XPB和XPD的相互作用可使UV诱导DNA损伤的宿主修复能力产生缺陷[25-28]。本研究结果显示，与HepG2细胞系比较，ETV-r HBV稳定转染后的HepG2.A64细胞系中RAD52和XRCC2基因mRNA和蛋白表达水平均显著降低。RAD52基因对维持肿瘤基因组的完整性至关重要，可在同源重组通路中驱动双链断裂修复时以RNA为模板的DNA反向链交换[29-30]，且RAD52中的SNPs与乳腺癌、肺癌、甲状腺癌、头颈癌和卵巢癌等多种癌症的发生风险有关[31]。此外，Sotiriou等[32]研究发现，RAD52与癌基因诱导的DNA复制应激反应有关。XRCC2基因同样为同源重组通路中的关键基因，参与将RAD51募集至DNA损伤部位，可以结合DNA并促进其同源配对[33]；有研究表明XRCC2突变的细胞具有更高的自发染色体不稳定水平，且对DNA链间交联剂(如丝裂霉素C和顺铂)、电离辐射以及烷基化剂和醛类表现出超敏性[34]，同时该基因的有害杂合突变会增加患乳腺癌的风险[35]。
综上所述，ETV-r HBV稳定转染后宿主细胞中出现大量差异表达基因，且宿主细胞同源重组修复中的关键基因RAD52及XRCC2表达量下调，提示HBV感染宿主细胞后可能通过劫持RAD52和XRCC2基因抑制宿主细胞的DNA修复能力，使受损DNA因不能得到及时有效的修复而不断累积，进而导致染色体不稳定、病毒基因组整合以及肿瘤发生。然而有关HBV感染抑制DNA损伤修复中关键基因的具体作用机制仍有待进一步研究。
基金
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  • 国家自然科学基金(81573676)
  • 北京市自然科学基金(7194302)
文献
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参考文献 引证文献
排序方式：
[1]
Yuen MF, Chen DS, Dusheiko GM, et al. Hepatitis B virus infection[J]. Nat Rev Dis Primers, 2018, 4: 18035.
[2]
Trépo C, Chan HLY, Lok A. Hepatitis B virus infection[J]. Lancet, 2014, 384(9959): 2053-2063.
[3]
Sells MA, Chen ML, Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA[J]. Proc Natl Acad Sci USA, 1987, 84(4): 1005-1009.
[4]
Ladner SK, Otto MJ, Barker CS, et al. Inducible expression of human hepatitis B virus (HBV) in stably transfected hepatoblastoma cells: a novel system for screening potential inhibitors of HBV replication[J]. Antimicrob Agents Chemother, 1997, 41(8): 1715-1720.
[5]
Sun DX, Nassal M. Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus[J]. J Hepatol, 2006, 45(5): 636-645.
[6]
Liu W, Song H, Chen Q, et al. Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of nucleos(t)ide analogues[J]. Liver Int, 2016, 36(9): 1284-1294.
[7]
Liu YT, Liu W, Zhu GY, et al. Involvement of multidrug resistance protein 4 in the hepatocyte efflux of lamivudine and entecavir[J]. Mol Med Rep, 2018, 17(5): 7113-7121.
[8]
Liu Y, Yao W, Si L, et al. Chinese herbal extract Su-duxing had potent inhibitory effects on both wild-type and entecavir-resistant hepatitis B virus (HBV) in vitro and effectively suppressed HBV replication in mouse model[J]. Antiviral Res, 2018, 155: 39-47.
[9]
Li H, Sheng C, Liu H, et al. An effective molecular target site in hepatitis B virus S gene for Cas9 cleavage and mutational inactivation[J]. Int J Biol Sci, 2016, 12(9): 1104-1113.
[10]
Li H, Sheng C, Wang S, et al. Removal of integrated hepatitis B virus DNA using CRISPR-Cas9[J]. Front Cell Infect Microbiol, 2017, 7: 91.
[11]
Ge LL, Yang XA, Piao JY, et al. Screening of tumor metastasis related differentially expressed genes in hepatocellular carcinoma cells with FATE/BJ-HCC-2 gene stable transfection by gene chip[J]. Chongqing Med, 2016, 45(12): 1595-1597.
[葛丽丽, 杨小昂, 朴军颜, 等. 基因芯片筛选稳定转染FATE/BJ-HCC-2基因的肝癌细胞中肿瘤转移相关差异表达基因[J]. 重庆医学, 2016, 45(12): 1595-1597.]
[12]
Wu Z, Wang T, Fang M, et al. MFAP5 promotes tumor progression and bone metastasis by regulating ERK/MMP signaling pathways in breast cancer[J]. Biochem Biophys Res Commun, 2018, 498(3): 495-501.
[13]
Li QM, Zhang YQ, Jiang QL. MFAP5 suppression inhibits migration/invasion, regulates cell cycle and induces apoptosis via promoting ROS production in cervical cancer[J]. Biochem Biophys Res Commun, 2018, 507(1/2/3/4): 51-58.
[14]
Vaittinen M, Kolehmainen M, Rydén M, et al. MFAP5 is related to obesity-associated adipose tissue and extracellular matrix remodeling and inflammation[J]. Obesity (Silver Spring), 2015, 23(7): 1371-1378.
[15]
Cheng Y, Wang KY, Geng LL, et al. Identification of candidate diagnostic and prognostic biomarkers for pancreatic carcinoma[J]. EBioMedicine, 2019, 40: 382-393.
[16]
Yang SF, Yeh CB, Chou YE, et al. Serpin peptidase inhibitor(SERPINB5) haplotypes are associated with susceptibility to hepatocellular carcinoma[J]. Sci Rep, 2016, 6: 26605.
[17]
Murphy CM, Xu YP, Li F, et al. Hepatitis B virus X protein promotes degradation of SMC5/6 to enhance HBV replication[J]. Cell Rep, 2016, 16(11): 2846-2854.
[18]
Decorsière A, Mueller H, van Breugel PC, et al. Hepatitis B virus X protein identifies the Smc5/6 complex as a host restriction factor[J]. Nature, 2016, 531(7594): 386-389.
[19]
Uhlmann F. SMC complexes: from DNA to chromosomes[J]. Nat Rev Mol Cell Biol, 2016, 17(7): 399-412.
[20]
Zhang Z, Huo H, Liao K, et al. RPA1 downregulation enhances nasopharyngeal cancer radiosensitivity via blocking RAD51 to the DNA damage site[J]. Exp Cell Res, 2018, 371(2): 330-341.
[21]
Sekiba K, Otsuka M, Ohno M, et al. Inhibition of HBV transcription from cccDNA With nitazoxanide by targeting the HBx-DDB1 interaction[J]. Cell Mol Gastroenterol Hepatol, 2019, 7(2): 297-312.
[22]
Gerossier L, Decorsière A, Abdul F, et al. A new restriction factor against HBV infection: the SMC5/6 complex[J]. Med Sci(Paris), 2016, 32(10): 808-812.
[23]
Rivière L, Quioc-Salomon B, Fallot G, et al. Hepatitis B virus replicating in hepatocellular carcinoma encodes HBx variants with preserved ability to antagonize restriction by Smc5/6[J]. Antiviral Res, 2019, 172: 104618.
[24]
Krupa R, Czarny P, Wigner P, et al. The relationship between single-nucleotide polymorphisms, the expression of DNA damage response genes, and hepatocellular carcinoma in a Polish population[J]. DNA Cell Biol, 2017, 36(8): 693-708.
[25]
Jaitovich-Groisman I, Benlimame N, Slagle BL, et al. Transcriptional regulation of the TFIIH transcription repair components XPB and XPD by the hepatitis B virus x protein in liver cells and transgenic liver tissue[J]. J Biol Chem, 2001, 276(17): 14124-14132.
[26]
Levrero M, Zucman-Rossi J. Mechanisms of HBV-induced hepatocellular carcinoma[J]. J Hepatol, 2016, 64(1 Suppl): S84-S101.
[27]
Qadri I, Fatima K, AbdeL-Hafiz H. Hepatitis B virus X protein impedes the DNA repair via its association with transcription factor, TFIIH[J]. BMC Microbiol, 2011, 11: 48.
[28]
Kokic G, Chernev A, Tegunov D, et al. Structural basis of TFIIH activation for nucleotide excision repair[J]. Nat Commun, 2019, 10(1): 2885.
[29]
Mazina OM, Keskin H, Hanamshet K, et al. Rad52 inverse strand exchange drives RNA-templated DNA double-strand break repair[J]. Mol Cell, 2017, 67(1): 19-29.e3.
[30]
Nogueira A, Fernandes M, Catarino R, et al. RAD52 functions in homologous recombination and its importance on genomic integrity maintenance and cancer therapy[J]. Cancers (Basel), 2019, 11(11): 1622.
[31]
Li HM, Yuan P, Yu DK, et al. Genetic variation in DNA repair gene RAD52 is associated with the response to platinum-based chemotherapy in SCLC patients[J]. Chin J Oncol, 2016, 38(7): 504-509.
[32]
Sotiriou SK, Kamileri I, Lugli N, et al. Mammalian RAD52 functions in break-induced replication repair of collapsed DNA replication forks[J]. Mol Cell, 2016, 64(6): 1127-1134.
[33]
Andreassen PR, Hanenberg H. XRCC2 (X-ray repair cross complementing 2)[J]. Atlas Genet Cytogenet Oncol Haematol, 2019, 23(1): 1-7.
[34]
Park JY, Virts EL, Jankowska A, et al. Complementation of hypersensitivity to DNA interstrand crosslinking agents demonstrates that XRCC2 is a Fanconi anaemia gene[J]. J Med Genet, 2016, 53(10): 672-680.
[35]
Hilbers FS, Luijsterburg MS, Wiegant WW, et al. Functional analysis of missense variants in the putative breast cancer susceptibility gene XRCC2[J]. Hum Mutat, 2016, 37(9): 914-925.
2021年第46卷第5期
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doi: 10.11855/j.issn.0577-7402.2021.05.04
  • 接收时间：2021-02-18
  • 首发时间：2025-12-24
  • 出版时间：2021-05-28
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  • 收稿日期：2021-02-18
  • 修回日期：2021-04-15
基金
National Natural Science Foundation of China(81573676)
国家自然科学基金(81573676)
Natural Science Foundation of Beijing(7194302)
北京市自然科学基金(7194302)
作者信息
    1病原微生物生物安全国家重点实验室，军事科学院军事医学研究院微生物流行病研究所，北京 100071
    2解放军总医院第五医学中心感染病医学部，北京 100039

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李浩，E-mail：;
刘妍，E-mail：
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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