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Article(id=1195362266108445466, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195362264082592240, articleNumber=1001-2494(2025)08-0818-10, orderNo=null, doi=10.11669/cpj.2025.08.006, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1736265600000, receivedDateStr=2025-01-08, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1762926172996, onlineDateStr=2025-11-12, pubDate=1744646400000, pubDateStr=2025-04-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1762926172996, onlineIssueDateStr=2025-11-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1762926172996, creator=13701087609, updateTime=1762926172996, updator=13701087609, issue=Issue{id=1195362264082592240, tenantId=1146029695717560320, journalId=1190317699101192196, year='2025', volume='60', issue='8', pageStart='777', pageEnd='890', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1762926172514, creator=13701087609, updateTime=1762928092119, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1195370315556635165, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195362264082592240, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1195370315560829470, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195362264082592240, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=818, endPage=827, ext={EN=ArticleExt(id=1195362266297189148, articleId=1195362266108445466, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Effect of Phellopterin on Preventing Breast Cancer Metastasis, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To investigate the role and mechanism of phellopterin in preventing breast cancer metastasis. METHODS The cytotoxicity of phellopterin was evaluated by CCK-8 assay and flow cytometry for cell cycle and apoptosis. The effect of phellopterin on the adhesion between cancer cells and vascular endothelial cells was detected by immunofluorescence. The effect of phellopterin on the expression of cell adhesion molecules (CAMs) on the surface of HUVECs was detected by flow cytometry. The effect of phellopterin on the expression of epithelial-mesenchymal transition(EMT) regulatory proteins in cancer cells was detected by Western blot and quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR). The inhibitory effect of phellopterin on lung metastasis of breast cancer in mice was evaluated by a mouse cancer metastasis model. RESULTS Phellopterin exhibits relatively low cytotoxicity and can inhibit the adhesion of cancer cells to HUVECs by reducing the expression of CAMs, such as vascular cell adhesion molecule-1(VCAM-1), intercellular cell adhesion molecule-1(ICAM-1), and E-selectin, which are induced by the inflammatory factor TNF-α. Additionally, phellopterin suppresses the expression of regulatory factors associated with EMT transformation and metastasis-promoting proteins, including tripartite motif containing 22(TRIM22), thrombospondin-1(THBS1), and KIT ligand(KITLG). In a mouse model of cancer metastasis, phellopterin was shown to significantly inhibit breast cancer lung metastasis and enhance the activity of anti-tumor immune cells (cytotoxic T lymphocytes and natural killer cells) in peripheral blood. CONCLUSION Phellopterin shows a significant preventive effect on breast cancer metastasis and holds substantial potential for further development and application.

, correspAuthors=Mingqing HUANG, Yusheng LU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Yinyin YAO, Zhouzhou YE, Weiyu WANG, Mengting LIN, Mingqing HUANG, Yusheng LU), CN=ArticleExt(id=1195362765176095160, articleId=1195362266108445466, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=珊瑚菜素预防乳腺癌转移的作用研究, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 研究珊瑚菜素在预防乳腺癌转移中的作用及机制。方法 通过CCK-8法、流式细胞术分析细胞周期和凋亡检测珊瑚菜素的细胞毒性,免疫荧光法检测珊瑚菜素对肿瘤细胞和血管内皮细胞之间黏附的影响,流式细胞术检测人静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)表面黏附分子表达的影响,Western blot和实时荧光定量聚合酶链式反应(qRT-PCR)法检测珊瑚菜素对肿瘤细胞上皮间充质转化(epithelial-mesenchymal transition,EMT)调控蛋白表达的影响,小鼠肿瘤转移模型评估珊瑚菜素对小鼠乳腺癌肺转移的抑制作用。结果 珊瑚菜素具有较低的细胞毒性,且在几乎无细胞毒性浓度下能够通过降低炎症因子肿瘤坏死因子(tumor necrosis factor α,TNF-α)诱导的HUVECs细胞表面血管细胞黏附分子-1(vascular cell adhesion molecule-1,CAM-1)、细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)和E-选择素(E-selectin)的表达;抑制肿瘤细胞在HUVECs上的黏附;抑制EMT转化相关调控因子及肿瘤细胞转移促进蛋白三联体基序蛋白22(tripartite motif protein 22,TRIM22)、血小板反应蛋白-1(thrombospondin-1,THBS1)和蛋白酪氨酸激酶KIT配体(KIT ligand,KITLG)的表达。小鼠肿瘤转移模型证明珊瑚菜素能够显著抑制乳腺癌肺转移,并激活小鼠外周血中抗肿瘤免疫细胞(细胞毒性T淋巴细胞和自然杀伤细胞)的活性。结论 珊瑚菜素展现出较好的预防乳腺癌转移作用,具有进一步开发应用价值。

, correspAuthors=黄鸣清, 卢余盛, authorNote=null, correspAuthorsNote=
*黄鸣清,男,博士,教授,博士生导师 研究方向:中药学 Tel:(0591)22861283;
卢余盛,男,博士,副研究员,硕士生导师 研究方向:抗肿瘤药物药 Tel:(0591)83761795
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姚茵茵,女,硕士研究生 研究方向:中药药理学

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姚茵茵,女,硕士研究生 研究方向:中药药理学

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DOI: 10.1186/s13046-018-0850-z., articleTitle=Hippo component YAP promotes focal adhesion and tumour aggressiveness via transcriptionally activating THBS1/FAK signalling in breast cancer, refAbstract=null), Reference(id=1195391152221373124, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, doi=null, pmid=null, pmcid=null, year=2019, volume=19, issue=1, pageStart=1015, pageEnd=null, url=null, language=null, rfNumber=[23], rfOrder=22, authorNames=XIAO H, HE M, XIE G, journalName=BMC Cancer, refType=null, unstructuredReference=XIAO H, HE M, XIE G, et al. The release of tryptase from mast cells promote tumor cell metastasis via exosomes[J]. BMC Cancer, 2019, 19(1):1015. DOI: 10.1186/s12885-019-6203-2., articleTitle=The release of tryptase from mast cells promote tumor cell metastasis via exosomes, refAbstract=null), Reference(id=1195391152284287685, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, doi=null, pmid=null, pmcid=null, year=2022, volume=102, issue=11, pageStart=1257, pageEnd=1267, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=LING J, ZHANG L, CHANG A, journalName=Lab Invest, refType=null, unstructuredReference=LING J, ZHANG L, CHANG A, et al. Overexpression of KITLG predicts unfavorable clinical outcomes and promotes lymph node metastasis via the JAK/STAT pathway in nasopharyngeal carcinoma[J]. Lab Invest, 2022, 102(11):1257-1267., articleTitle=Overexpression of KITLG predicts unfavorable clinical outcomes and promotes lymph node metastasis via the JAK/STAT pathway in nasopharyngeal carcinoma, refAbstract=null)], funds=[Fund(id=1195391150455571116, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, awardId=81703555, language=CN, fundingSource=国家自然科学基金项目资助(81703555), fundOrder=null, country=null), Fund(id=1195391150522679981, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, awardId=2020J01859, language=CN, fundingSource=福建省自然科学基金项目资助(2020J01859), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1195391146491953763, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, xref=1, ext=[AuthorCompanyExt(id=1195391146500342372, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, companyId=1195391146491953763, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 Fujian Key Laboratory of Chinese Materia Medica, College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122 China), AuthorCompanyExt(id=1195391146508730981, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, companyId=1195391146491953763, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 福建中医药大学药学院, 福建省中药学重点实验室, 福州 350122)]), AuthorCompany(id=1195391146567451238, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, xref=2, ext=[AuthorCompanyExt(id=1195391146571645543, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, companyId=1195391146567451238, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Fuzhou Institute of Oceanography, College of Material and Chemical Engineering, Minjiang University, Fuzhou 350108, China), AuthorCompanyExt(id=1195391146580034152, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, companyId=1195391146567451238, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 福州海洋研究院, 闽江学院材料与化学工程学院, 福州 350108)])], figs=[ArticleFig(id=1195391148954010262, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.1, caption=Effects of phellopterin (PHE) on the viability of MCF-7 and 4T1 cells.n=6,$\stackrel{-}{x}$±s, figureFileSmall=8xGSZF/SQiURlQoff/3lrA==, figureFileBig=KxbWNz1IyjQc7pXjdfg+NA==, tableContent=null), ArticleFig(id=1195391149021119127, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图1, caption=珊瑚菜素对人乳腺癌MCF-7细胞和小鼠乳腺癌4T1细胞的活性影响。n=6,$\stackrel{-}{x}$±s, figureFileSmall=8xGSZF/SQiURlQoff/3lrA==, figureFileBig=KxbWNz1IyjQc7pXjdfg+NA==, tableContent=null), ArticleFig(id=1195391149134365336, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.2, caption=The effects of PHE on apoptosis and cell cycle distribution in HUVECs (panels A and C) and MCF-7 cells (panels B and D), figureFileSmall=I4dhsiERp3IS8LIkg1HzJw==, figureFileBig=ucfFMAA2e5fimXDLnymxOw==, tableContent=null), ArticleFig(id=1195391149193085593, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图2, caption=珊瑚菜素对HUVECs(A和C)和MCF-7(B和D)细胞凋亡和细胞周期分布的影响, figureFileSmall=I4dhsiERp3IS8LIkg1HzJw==, figureFileBig=ucfFMAA2e5fimXDLnymxOw==, tableContent=null), ArticleFig(id=1195391149256000154, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.3, caption=Effects of PHE on cell adhesion and the expression of adhesion molecules.n=3,$\stackrel{-}{x}$±s

A-fluorescence microscopy images show MCF-7 cells (labeled with rhodamine 123, shown in green) adhering to HUVECs; B-PHE significantly inhibits the adhesion of MCF-7 cells to HUVECs induced by the inflammatory factor TNF-α; C-E-flow cytometry was used to detect the expression of adhesion molecules VCAM-1 (CD106), E-selectin (CD62E), and ICAM-1 (CD54) on the surface of HUVECs; F-H-PHE significantly inhibits the expression of cell adhesion molecules(CAMs) on the surface of TNF-α-activated HUVECs;1)P<0.05, 2)P<0.01, 3)P<0.001, compared with the control (TNF-α treatment only) group.

, figureFileSmall=5v4KZaTJRHf6B+9zqPA7aw==, figureFileBig=3s1Xgujfz76bfqWTh4bh/w==, tableContent=null), ArticleFig(id=1195391149323109019, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图3, caption=珊瑚菜素对细胞黏附和黏附分子表达的影响。n=3,$\stackrel{-}{x}$±s

A-荧光显微镜成像显示MCF-7细胞(罗丹明123标记,绿色)黏附在HUVECs细胞上;B-珊瑚菜素显著抑制炎症因子肿瘤坏死因子(tumor necrosis factor-α,TNF-α)诱导的MCF-7细胞在HUVECs上的黏附;C~E-流式细胞术检测HUVECs表面黏附分子血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1,CD106)、细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1,CD54)和E-选择素(E-selectin,CD62E)的表达;F~H-珊瑚菜素显著抑制TNF-α激活的HUVECs表面黏附分子的表达;与对照组(仅TNF-α治疗)比较,1)P<0.05,2)P<0.01,3)P<0.001。

, figureFileSmall=5v4KZaTJRHf6B+9zqPA7aw==, figureFileBig=3s1Xgujfz76bfqWTh4bh/w==, tableContent=null), ArticleFig(id=1195391149381829276, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.4, caption=Effects of PHE on the expression of regulatory factors of epithelial mesenchymal transformation (EMT) was detected by quantitative PCR. n=3,$\stackrel{-}{x}$±s

1)P<0.05, 2)P<0.01, 3)P<0.001, compared with the control group.

, figureFileSmall=qf68oIj1C+22vdUbq3WKMw==, figureFileBig=6/zwUQREHtLkFvq1Ltot4A==, tableContent=null), ArticleFig(id=1195391149440549533, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图4, caption=qRT-PCR检测珊瑚菜素对上皮间充质转化调控因子表达的影响。n=3,$\stackrel{-}{x}$±s

与对照组比较,1)P<0.05,2)P<0.01,3)P<0.001。

, figureFileSmall=qf68oIj1C+22vdUbq3WKMw==, figureFileBig=6/zwUQREHtLkFvq1Ltot4A==, tableContent=null), ArticleFig(id=1195391149503464094, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.5, caption=Effects of PHE on the expression of metastasis-promoting proteins TRIM22, THBS1 and KITLG were detected by Western blot (A-D) and quantitative PCR (E-G).n=3,$\stackrel{-}{x}$±s

1)P<0.05, 2)P<0.01, compared with the control group.

, figureFileSmall=ysJ6aecx3SXsmrDI6hkP3g==, figureFileBig=2DHyF5KkNBdLxsLmMcLxkg==, tableContent=null), ArticleFig(id=1195391149557990047, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图5, caption=Western blot(A~D)和qRT-PCR(E~G)检测珊瑚菜素对促转移蛋白三联基元22(TRIM22)、血小板反应素-1(THBS1)和重组人干细胞因子(KITLG)表达的影响。n=3,$\stackrel{-}{x}$±s

与对照组比较,1)P<0.05,2)P<0.01。

, figureFileSmall=ysJ6aecx3SXsmrDI6hkP3g==, figureFileBig=2DHyF5KkNBdLxsLmMcLxkg==, tableContent=null), ArticleFig(id=1195391149616710304, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.6, caption=Effects of PHE on lung metastasis of breast cancer in mice was evaluated by a mouse cancer metastasis model. n=6,$\stackrel{-}{x}$±s

A-the formation of lung cancer nodules was observed in vivo in mice; B-C-PHE concentration-dependently reduced the number of lung metastatic nodules (B) and lung mass (C) in mice; 1)P<0.05, compared with the control group (0 mg·kg-1).

, figureFileSmall=7tyh3vyYWKhu1GDauLwWcQ==, figureFileBig=O9m8GfIpiV3tSf2vkKte0g==, tableContent=null), ArticleFig(id=1195391149683819169, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图6, caption=小鼠肿瘤转移模型评估珊瑚菜素对小鼠乳腺癌肺转移的影响。n=6,$\stackrel{-}{x}$±s

A-小鼠肺组织解剖图;B~C-珊瑚菜素浓度依赖的降低小鼠肺部乳腺癌转移结节数(B)和肺质量(C);与对照组(0 mg·kg-1)比较,1)P<0.05。

, figureFileSmall=7tyh3vyYWKhu1GDauLwWcQ==, figureFileBig=O9m8GfIpiV3tSf2vkKte0g==, tableContent=null), ArticleFig(id=1195391149746733730, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.7, caption=Images of lung, liver, spleen, kidney and heart sections stained with hematoxylin-eosin (HE) from PHE-treated mice

The black arrows in the lung section indicate the area of breast cancer metastasis.

, figureFileSmall=a0mR9Mcs+PhlyPAL15CyQA==, figureFileBig=8Wk5lK2F/6KdO6yV6eC2Tw==, tableContent=null), ArticleFig(id=1195391149813842595, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图7, caption=经珊瑚菜素治疗的小鼠肺、肝、脾、肾和心脏切片的苏木精-伊红(HE)染色代表图

肺切片中黑色箭头指示为乳腺癌转移的区域。

, figureFileSmall=a0mR9Mcs+PhlyPAL15CyQA==, figureFileBig=8Wk5lK2F/6KdO6yV6eC2Tw==, tableContent=null), ArticleFig(id=1195391149876757156, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.8, caption=Quantitative statistical map of pulmonary nodules in mouse HE staining. n=6,$\stackrel{-}{x}$±s

1)P<0.05, compared with the control group.

, figureFileSmall=gMF1jqXbdBbShIgKFWoD9w==, figureFileBig=xfVKyYiJ8pxK+FaP6PQ2Xw==, tableContent=null), ArticleFig(id=1195391149943866021, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图8, caption=小鼠HE染色中肺结节的量化统计图。n=6,$\stackrel{-}{x}$±s

与对照组比较,1)P<0.05。

, figureFileSmall=gMF1jqXbdBbShIgKFWoD9w==, figureFileBig=xfVKyYiJ8pxK+FaP6PQ2Xw==, tableContent=null), ArticleFig(id=1195391150010974886, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.9, caption=Quantification of cytotoxic T lymphocytes(Tc,CD3+CD8+)cells percentage in mice peripheral blood by flow cytometry. n=6,$\stackrel{-}{x}$±s

1)P<0.05, 2)P<0.01, compared with the control group(0 mg·kg-1).

, figureFileSmall=HTnuR5+JwyEbEXecFb4gow==, figureFileBig=yJ+GpRF5oTKq9xZJrqhBtA==, tableContent=null), ArticleFig(id=1195391150073889447, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图9, caption=流式细胞术检测小鼠外周血中细胞毒性T细胞(Tc细胞,CD3+CD8+)百分含量。n=6,$\stackrel{-}{x}$±s

与对照组(0 mg·kg-1)比较,1)P<0.05,2)P<0.01。

, figureFileSmall=HTnuR5+JwyEbEXecFb4gow==, figureFileBig=yJ+GpRF5oTKq9xZJrqhBtA==, tableContent=null), ArticleFig(id=1195391150132609704, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Fig.10, caption=Quantification of natural killer(NK,CD49+)cells percentage in mice peripheral blood by flow cytometry. n=6,$\stackrel{-}{x}$±s

1)P<0.05,2)P<0.01, compared to the control (0 mg·kg-1).

, figureFileSmall=6cBbKSxfgroe9vYsd5s2rA==, figureFileBig=4N94zZqwp1A0yDIU+5r14g==, tableContent=null), ArticleFig(id=1195391150203912873, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=图10, caption=流式细胞术检测小鼠外周血中自然杀伤细胞(NK细胞,CD49+)百分含量。n=6,$\stackrel{-}{x}$±s

与对照组(0 mg·kg-1)比较,1)P<0.05,2)P<0.01。

, figureFileSmall=6cBbKSxfgroe9vYsd5s2rA==, figureFileBig=4N94zZqwp1A0yDIU+5r14g==, tableContent=null), ArticleFig(id=1195391150262633130, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=EN, label=Tab.1, caption=

qRT-PCR primers.

, figureFileSmall=null, figureFileBig=null, tableContent=
Primers Primer sequence
Forward primer Reverse primer
THBS1 5'-AGTCGTCTCTGCAACAACCC-3' 5'-GGGATTGGACAGGCATCCAT-3'
KITLG 5'-CCGGGATGGATGTTTTGCCA-3' 5'-GCACTCCACAAGGTCATCCA-3'
MMP2 5'-GGCATTCAGGAGCTCTATGG-3' 5'-GAGCGATGCCATCAAATACA-3'
HIF1 5'-CGTTCCTTCGATCAGTTGTC-3' 5'-TCAGTGGTGGCAGTGGTAGT-3'
CDH2 5'-GACCCAAACAGCAACGACGGGT-3' 5'-CTGAGGCGGGTGCTGAATTCCC-3'
ZEB2 5'-ACGGTATTGCCAACCCTCTG-3' 5'-CTCCCTTATTTCATCTTCCTCTTC-3'
ZEB1 5'-ATCCTGGGGCCTGAAGCTCAGG-3' 5'-TGGTGTGCCCTGCCTCTGGT-3'
SNAIL1 5'-GCTGCTACAAGGCCATGTCCGG-3' 5'-CTTGGTGCTTGTGGAGCAGGGAC-3'
SNAIL2 5'-GATGCCGCGCTCCTTCCTGG-3' 5'-TGGAGCAGCGGTAGTCCACAC-3'
FN1 5'-TGCAAGGCCTCAGACCGGGT-3' 5'-GCGCTCAGGCTTGTGGGTGT-3'
TWIST 5'-TCCGCGTCCCACTAGCAGGC-3' 5'-CGCCCCACGCCCTGTTTCTT-3'
GAPDH 5'-GTCAAGGCTGAGAACGGGAA-3' 5'-AAATGAGCCCCAGCCTTCTC-3'
TRIM22 5'-TGAGTGGGATGCTGCAAGTT-3' 5'-CAGCCGAAGACACCAAAAGC-3'
), ArticleFig(id=1195391150325547691, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195362266108445466, language=CN, label=表1, caption=

实时荧光定量聚合酶链式反应(qRT-PCR)引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
Primers Primer sequence
Forward primer Reverse primer
THBS1 5'-AGTCGTCTCTGCAACAACCC-3' 5'-GGGATTGGACAGGCATCCAT-3'
KITLG 5'-CCGGGATGGATGTTTTGCCA-3' 5'-GCACTCCACAAGGTCATCCA-3'
MMP2 5'-GGCATTCAGGAGCTCTATGG-3' 5'-GAGCGATGCCATCAAATACA-3'
HIF1 5'-CGTTCCTTCGATCAGTTGTC-3' 5'-TCAGTGGTGGCAGTGGTAGT-3'
CDH2 5'-GACCCAAACAGCAACGACGGGT-3' 5'-CTGAGGCGGGTGCTGAATTCCC-3'
ZEB2 5'-ACGGTATTGCCAACCCTCTG-3' 5'-CTCCCTTATTTCATCTTCCTCTTC-3'
ZEB1 5'-ATCCTGGGGCCTGAAGCTCAGG-3' 5'-TGGTGTGCCCTGCCTCTGGT-3'
SNAIL1 5'-GCTGCTACAAGGCCATGTCCGG-3' 5'-CTTGGTGCTTGTGGAGCAGGGAC-3'
SNAIL2 5'-GATGCCGCGCTCCTTCCTGG-3' 5'-TGGAGCAGCGGTAGTCCACAC-3'
FN1 5'-TGCAAGGCCTCAGACCGGGT-3' 5'-GCGCTCAGGCTTGTGGGTGT-3'
TWIST 5'-TCCGCGTCCCACTAGCAGGC-3' 5'-CGCCCCACGCCCTGTTTCTT-3'
GAPDH 5'-GTCAAGGCTGAGAACGGGAA-3' 5'-AAATGAGCCCCAGCCTTCTC-3'
TRIM22 5'-TGAGTGGGATGCTGCAAGTT-3' 5'-CAGCCGAAGACACCAAAAGC-3'
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珊瑚菜素预防乳腺癌转移的作用研究
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姚茵茵 1, 2 , 叶周周 2 , 王伟雨 1, 2 , 林梦婷 1, 2 , 黄鸣清 1, * , 卢余盛 2, *
中国药学杂志 | 论著 2025,60(8): 818-827
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中国药学杂志 | 论著 2025, 60(8): 818-827
珊瑚菜素预防乳腺癌转移的作用研究
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姚茵茵1, 2, 叶周周2, 王伟雨1, 2, 林梦婷1, 2, 黄鸣清1, *, 卢余盛2, *
作者信息
  • 1 福建中医药大学药学院, 福建省中药学重点实验室, 福州 350122
  • 2 福州海洋研究院, 闽江学院材料与化学工程学院, 福州 350108

    • 姚茵茵,女,硕士研究生 研究方向:中药药理学

通讯作者:

*黄鸣清,男,博士,教授,博士生导师 研究方向:中药学 Tel:(0591)22861283;
卢余盛,男,博士,副研究员,硕士生导师 研究方向:抗肿瘤药物药 Tel:(0591)83761795
Effect of Phellopterin on Preventing Breast Cancer Metastasis
Yinyin YAO1, 2, Zhouzhou YE2, Weiyu WANG1, 2, Mengting LIN1, 2, Mingqing HUANG1, *, Yusheng LU2, *
Affiliations
  • 1 Fujian Key Laboratory of Chinese Materia Medica, College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122 China
  • 2 Fuzhou Institute of Oceanography, College of Material and Chemical Engineering, Minjiang University, Fuzhou 350108, China
出版时间: 2025-04-15 doi: 10.11669/cpj.2025.08.006
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摘要
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目的 研究珊瑚菜素在预防乳腺癌转移中的作用及机制。方法 通过CCK-8法、流式细胞术分析细胞周期和凋亡检测珊瑚菜素的细胞毒性,免疫荧光法检测珊瑚菜素对肿瘤细胞和血管内皮细胞之间黏附的影响,流式细胞术检测人静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)表面黏附分子表达的影响,Western blot和实时荧光定量聚合酶链式反应(qRT-PCR)法检测珊瑚菜素对肿瘤细胞上皮间充质转化(epithelial-mesenchymal transition,EMT)调控蛋白表达的影响,小鼠肿瘤转移模型评估珊瑚菜素对小鼠乳腺癌肺转移的抑制作用。结果 珊瑚菜素具有较低的细胞毒性,且在几乎无细胞毒性浓度下能够通过降低炎症因子肿瘤坏死因子(tumor necrosis factor α,TNF-α)诱导的HUVECs细胞表面血管细胞黏附分子-1(vascular cell adhesion molecule-1,CAM-1)、细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)和E-选择素(E-selectin)的表达;抑制肿瘤细胞在HUVECs上的黏附;抑制EMT转化相关调控因子及肿瘤细胞转移促进蛋白三联体基序蛋白22(tripartite motif protein 22,TRIM22)、血小板反应蛋白-1(thrombospondin-1,THBS1)和蛋白酪氨酸激酶KIT配体(KIT ligand,KITLG)的表达。小鼠肿瘤转移模型证明珊瑚菜素能够显著抑制乳腺癌肺转移,并激活小鼠外周血中抗肿瘤免疫细胞(细胞毒性T淋巴细胞和自然杀伤细胞)的活性。结论 珊瑚菜素展现出较好的预防乳腺癌转移作用,具有进一步开发应用价值。

珊瑚菜素  /  肿瘤转移  /  细胞黏附分子  /  上皮间充质转化  /  免疫激活

OBJECTIVE To investigate the role and mechanism of phellopterin in preventing breast cancer metastasis. METHODS The cytotoxicity of phellopterin was evaluated by CCK-8 assay and flow cytometry for cell cycle and apoptosis. The effect of phellopterin on the adhesion between cancer cells and vascular endothelial cells was detected by immunofluorescence. The effect of phellopterin on the expression of cell adhesion molecules (CAMs) on the surface of HUVECs was detected by flow cytometry. The effect of phellopterin on the expression of epithelial-mesenchymal transition(EMT) regulatory proteins in cancer cells was detected by Western blot and quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR). The inhibitory effect of phellopterin on lung metastasis of breast cancer in mice was evaluated by a mouse cancer metastasis model. RESULTS Phellopterin exhibits relatively low cytotoxicity and can inhibit the adhesion of cancer cells to HUVECs by reducing the expression of CAMs, such as vascular cell adhesion molecule-1(VCAM-1), intercellular cell adhesion molecule-1(ICAM-1), and E-selectin, which are induced by the inflammatory factor TNF-α. Additionally, phellopterin suppresses the expression of regulatory factors associated with EMT transformation and metastasis-promoting proteins, including tripartite motif containing 22(TRIM22), thrombospondin-1(THBS1), and KIT ligand(KITLG). In a mouse model of cancer metastasis, phellopterin was shown to significantly inhibit breast cancer lung metastasis and enhance the activity of anti-tumor immune cells (cytotoxic T lymphocytes and natural killer cells) in peripheral blood. CONCLUSION Phellopterin shows a significant preventive effect on breast cancer metastasis and holds substantial potential for further development and application.

phellopterin  /  cancer metastasis  /  cell adhesion molecule  /  epithelial-mesenchymal transition  /  immune activation
姚茵茵, 叶周周, 王伟雨, 林梦婷, 黄鸣清, 卢余盛. 珊瑚菜素预防乳腺癌转移的作用研究. 中国药学杂志, 2025 , 60 (8) : 818 -827 . DOI: 10.11669/cpj.2025.08.006
Yinyin YAO, Zhouzhou YE, Weiyu WANG, Mengting LIN, Mingqing HUANG, Yusheng LU. Effect of Phellopterin on Preventing Breast Cancer Metastasis[J]. Chinese Pharmaceutical Journal, 2025 , 60 (8) : 818 -827 . DOI: 10.11669/cpj.2025.08.006
正文
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肿瘤的复发与转移是导致肿瘤治疗失败的主要原因,困扰着人数逐年增加的数以亿计的肿瘤幸存者[1]。当前,肿瘤治疗策略涵盖了放射治疗、化学疗法、免疫治疗及手术治疗等多元化手段[2-3],然而这些疗法仍难以遏制肿瘤的复发与转移。肿瘤转移是一个涉及多阶段的恶性过程,包括肿瘤细胞从原发部位脱落、穿过血管内皮进入血液循环形成循环肿瘤细胞(circulating tumor cells,CTCs)、在循环系统中逃避免疫攻击、特定器官内的黏附与跨内皮细胞迁移,以及在新器官内的增殖与血管生成等[4-6]。CTCs在血管内皮细胞上的黏附及随后的跨内皮细胞迁移是肿瘤形成远端转移的两个重要环节[7-8]。研究表明,血管内皮细胞表面细胞黏附分子(cell adhesion molecules,CAMs)能够促进肿瘤细胞的黏附[9],而肿瘤细胞上皮间充质转化(epithelial-mesenchymal transition,EMT)能够促进肿瘤转移过程[10]。因此,从干预肿瘤转移过程入手,发现能有效预防肿瘤转移的新化合物是当前肿瘤治疗关注的热点之一[11-13]。
珊瑚菜素是一种香豆素类化合物,主要来源于伞形科植物珊瑚菜(Glehnia littoralis F. Schmidt ex Miq.)、白芷[Angelica dahurica (Fisch. ex Hoffm.) Benth. & Hook. f. ex Franch. & Sav.]等,具有抗病毒、抗炎、抗肿瘤和增强免疫系统功能等多种药理药效,在中医药研究及药物开发领域展现出广阔的应用前景[14-16]。目前,尚未有关于珊瑚菜素在抑制CTCs黏附于血管内膜、跨内皮迁移及预防肿瘤转移方面的研究。本研究将通过体内外实验探讨珊瑚菜素在预防乳腺癌转移过程中的作用及其机制,进一步开发珊瑚菜素的潜在应用价值。
1 仪器与试剂
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1.1 仪器
多功能酶标仪(型号:M200 PRO,瑞士TECAN公司);荧光显微镜(型号:DMIL LED,德国LEICA公司);分选型流式细胞仪(型号:FACS AriaIII)、流式细胞仪(型号:FACSCelestaTM,美国BD公司);实时荧光定量聚合酶链式反应仪(qRT-PCR,型号:CFX96TM)、化学发光成像系统(型号:ChemiDocTMMP)(美国Bio-Rad公司)。
1.2 试剂
珊瑚菜素(货号:DS0110,成都德思特生物技术有限公司,纯度大于98%);杜氏改良Eagle培养基(DMEM,批号:SH30022.01,美国HyClone公司);内皮细胞培养基(ECM,批号:34840)、内皮细胞生长补充剂(ECGS,批号:29377)(美国ScienCell公司);肿瘤坏死因子-α(TNF-α,批号:8902SC,美国CST公司);罗丹明123(批号:6AS1012,鼎国昌盛生物技术有限公司);CCK-8试剂盒(批号:P1030)、Trizol(批号:ET111-01-V2)、PerfectStart Green qPCR SuperMix TransScriptAll-in-One First-Strand(批号:AQ601-04)、cDNA Synthesis SuperMix for qPCR(批号:AT341-02)(北京全式金生物技术有限公司);细胞周期检测试剂盒(批号:CA002,美国SAB公司);细胞凋亡检测试剂盒(批号:GK0603-50T)、磷酸盐缓冲液(PBS,批号:GS3101-500ML)(美国Genview公司);Anti-Human CD54-FITC抗体(批号:559771)、Anti-Human CD62E-APC抗体(批号:551144)、Anti-Human CD106-PE抗体(批号:2094221)(美国BD PharmingenTM公司);GAPDH抗体(批号:A19056)、血小板反应素蛋白1抗体(THBS1,批号:A2125)、蛋白酪氨酸激酶KIT配体(KITLG)抗体(批号:A16245)(武汉爱博泰克生物科技有限公司);三联体基序蛋白22 (TRIM22)抗体(批号:240411D6,武汉三鹰生物技术有限公司);HRP Goat Anti-Rabbit IgG(批号:GAR007)、Anti-Mouse CD3-PerCP-Cy5.5抗体(批号:AM003E07)、Anti-Mouse CD45-V450抗体(批号:AM04512)、Anti-Mouse CD8-PE-Cy7抗体(批号:M008A10)(杭州联科生物技术有限公司)。
1.3 细胞与实验动物
人静脉内皮细胞(HUVECs)细胞株为本实验室提取保存,人乳腺癌MCF-7细胞和小鼠乳腺癌4T1细胞株(中国科学院上海细胞库);SPF级别的BALB/c雌性小鼠,体质量(20±2)g,年龄6~8周(北京华阜康生物科技股份有限公司)[生产许可证:SCXK(京)2019-0008]。动物实验均根据《实验室动物护理和使用指南》(国家研究委员会,1996年)实行。闽江学院实验动物伦理委员会审核并批准了所有动物使用程序。
2 方法
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2.1 细胞实验
2.1.1 细胞培养
将HUVECs细胞和MCF-7复苏后,分别用含体积分数为20%胎牛血清(FBS)、1%双抗、1%ECGS的ECM培养基和体积分数10%FBS、2%双抗的DMEM培养基,培养在37 ℃,5%CO2的培养箱中。待细胞密度达到80%左右进行消化传代、冻存。取状态良好的细胞完成后续实验。
2.1.2 CCK-8检测珊瑚菜素对细胞活力的影响
分别将MCF-7细胞与4T1细胞以5 000个每孔的细胞密度接种于96孔板上培养过夜,分别加入珊瑚菜素物质的量浓度为0、1、5、10、50、100、200、500、1 000 μmol·L-1的培养液,每组6个复孔,培养24 h。除去含药培养液,加入体积分数10%的CCK-8工作液,孵育1~2 h后,多功能酶标仪在450 nm处测量吸光度值,计算细胞半数抑制浓度(IC50)。
2.1.3 细胞周期实验
取对数生长期的MCF-7细胞接种于6孔板中,待细胞密度达到60%左右,分别加入珊瑚菜素物质的量浓度为0、1、10、50、100 μmol·L-1的培养液,培养24 h。孵育结束收集细胞沉淀,用预冷体积分数75%的乙醇固定过夜;离心收集并清洗细胞沉淀后,加入20 μL核糖核酸酶A(RnaseA)溶液于37 ℃水浴30 min;加入400 μL碘化丙啶染液(PI)于4 ℃避光孵育30 min。随后采用流式细胞仪进行检测,FlowJo软件分析作图。
2.1.4 细胞凋亡实验
将MCF-7细胞接种于6孔板中,待细胞密度达到60%左右,加入珊瑚菜素浓度为0、1、10、50、100 μmol·L-1的培养液,培养24 h。孵育结束收集培养液与消化好的细胞悬液,离心取细胞沉淀,用400 μL 1×Annexin V结合液悬浮细胞,并加入5 μL Annexin V-FITC染色液和5 μL PI染液,混匀后室温避光孵育15 min。随后采用流式细胞仪进行检测,FlowJo软件分析作图。
2.1.5 珊瑚菜素对MCF-7细胞黏附于内皮细胞的影响
取3~6代状态良好的HUVECs细胞均匀铺于24孔板中,待细胞密度达到95%时,给予TNF-α(10 ng·mL-1)刺激4 h,另设置一个阴性对照组。配制含不同物质的量浓度(0、1、10、50、100 μmol·L-1)珊瑚菜素的培养液;消化收集MCF-7细胞,并用罗丹明染色;分别用配好的含珊瑚菜素培养液将染色的MCF-7细胞调整为每毫升5×105个;将细胞悬液分别加到TNF-α刺激4 h的HUVECs细胞中,继续培养1 h。孵育结束,除去未黏附细胞,使用荧光显微镜进行拍照并计数,计算黏附率。
2.1.6 珊瑚菜素对内皮细胞表面黏附分子表达的影响
HUVECs细胞均匀铺于6孔板中,待细胞密度达到60%时,加入含不同物质的量浓度(0、1、10、50、100 μmol·L-1)珊瑚菜素的培养基,放入培养箱继续培养;20 h后给予TNF-α(10 ng·mL-1)刺激4 h,另设置一个阴性对照组。消化收集细胞,用200 μL PBS重悬细胞,分别用CD54-APC、CD106-PE、CD62E-APC抗体进行染色,4 ℃避光孵育30 min;最后,收集并清洗细胞沉淀,用PBS重悬后,使用分选型流式细胞仪进行检测。
2.1.7 Western blot
将MCF-7细胞接种于6孔板中,待细胞密度达到60%左右,加入含不同物质的量浓度(分别为0、10、50、100 μmol·L-1)珊瑚菜素的培养液,培养箱培养24 h。用裂解液(蛋白酶和磷酸酶抑制剂混合液-RIPA=1∶100)分别裂解细胞,提取细胞总蛋白。将定量后的样品加到4%~20%梯度预制胶中,电泳分离、切胶、转膜;用1×Tris缓冲盐Tween-20缓冲液(TBST)清洗条带后,质量分数5%的脱脂奶粉封闭1 h;随后分别加入一抗GAPDH(1∶5 000)、THBS1(1∶2 000)、KITLG(1∶2 000)、TRIM22(1∶2 000)于4 ℃过夜;隔天清洗条带后,加入二抗(1∶10 000)孵育1 h;洗膜后加入高敏化学发光试剂(ECL)工作液,于化学发光成像仪显影,软件计算相对表达量。
2.1.8 实时荧光定量PCR(qRT-PCR)
细胞培养与Western blot所述相同,按试剂盒说明书用Trizol提取各组细胞的总RNA,接着将RNA逆转录成cDNA。在八联管加入各个组分混合均匀,放入qRT-PCR仪进行定量分析。PCR反应条件为:94 ℃预变性30 s;94 ℃,5 s;60 ℃,30 s,共循环40次;94 ℃,15 s;60 ℃,60 s;94 ℃,15 s。引物序列见表1。
2.2 动物实验
2.2.1 小鼠乳腺癌肺转移模型的建立与给药
BALB/c雌性小鼠随机分组,分别为正常组(Normal)、肿瘤组(Control)、肿瘤+珊瑚菜素低剂量组(1 mg·kg-1)、肿瘤+珊瑚菜素中剂量组(5 mg·kg-1)、肿瘤+珊瑚菜素高剂量组(10 mg·kg-1),每组6只。构建转移模型的前3 d通过腹腔注射给药治疗。消化收集4T1细胞,用PBS调整成每毫升5×105个的细胞悬液,通过尾静脉注射100 μL细胞悬液构建小鼠乳腺癌转移模型。造模后继续腹腔注射给予对应药物治疗21 d。
2.2.2 珊瑚菜素对小鼠肺部转移的影响
末次给药24 h后,小鼠摘眼球取血,脱颈处死。对小鼠进行解剖,取出肺称重后用生理盐水清洗干净,用Bouin’s固定液固定肺组织,观察肿瘤转移情况。其他组织用生理盐水清洗后,固定于质量分数为4%的多聚甲醛中。经脱水包埋、切片后,进行苏木精-伊红染色(hematoxylin-eosin staining,HE)法,结果用显微镜拍摄观察,HE染色中的肺结节大小使用Image J软件进行定量分析。
2.2.3 免疫细胞的检测
收集各组小鼠外周血于抗凝管中,并加入适量CD45-V450、CD3-PerCP-Cy5.5、CD8-PE-Cy7和CD49-FITC抗体于4 ℃避光孵育30 min;孵育结束后加入红细胞裂解液进行破红处理,在37 ℃水浴锅温浴10 min;随后离心收集细胞沉淀,并用PBS清洗、重悬,采用流式细胞仪检测各组免疫细胞的含量,FlowJo软件分析作图。
2.3 统计学分析
本研究数据使用 GraphPad Prism软件进行统计分析并作图。组间经单因素方差分析(ANOVA),各组数据平均值间差异采用邓尼特多重比较检验。当P<0.05时,表示组间统计学上有显著差异,具有统计学意义;当P<0.01时,表示组间统计学上有非常显著差异,具有统计学意义。
3 结果
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3.1 珊瑚菜素对乳腺癌细胞活性的影响
采用CCK-8法评估珊瑚菜素对MC-7和4T1细胞活性的影响,结果见图1,在珊瑚菜素物质的量浓度低于100 μmol·L-1时,MCF-7与4T1细胞的存活率都在90%以上。即使珊瑚菜素物质的量浓度达到500 μmol·L-1时,也没有达到细胞的IC50。由此可见,当珊瑚菜素的物质的量浓度低于100 μmol·L-1时没有明显的细胞毒性。
3.2 珊瑚菜素对HUVECs和MCF-7细胞周期和凋亡的影响
进一步采用流式细胞术检测HUVECs和MCF-7细胞凋亡与周期的影响。结果见图2,在物质的量浓度0~100 μmol·L-1的范围内,各组的正常细胞占比均在98%以上,没有明显的诱导细胞凋亡(图2A~B)。细胞周期实验同样表明,珊瑚菜素没有显著地抑制细胞分裂(图2C~D),说明珊瑚菜素物质的量浓度低于100 μmol·L-1时仅仅具有低的细胞毒性。因此,在后续的预防肿瘤细胞转移实验中,珊瑚菜素治疗浓度均低于100 μmol·L-1,以排除细胞毒性作用对实验结果的影响。
3.3 珊瑚菜素通过抑制黏附分子表达阻止MCF-7细胞在血管内皮细胞上的黏附
肿瘤细胞黏附于血管内膜是形成转移的关键步骤之一。本研究采用黏附实验测试珊瑚菜素对MCF-7细胞黏附于HUVECs细胞的影响。结果见图3A~B,炎症因子TNF-α能显著促进MCF-7在HUVECs上的黏附,随着PHE浓度升高,能显著抑制MCF-7的黏附。进一步采用流式细胞仪检测珊瑚菜素对HUVECs表面细胞间黏附分子-1(ICAM-1)(CD54)、血管细胞黏附分子-1(VCAM-1)(CD106)、E选择素(E-seletin)(CD62E)表达的影响,结果见图3C~H,炎症因子TNF-α能显著激活HUVECs表面黏附分子的表达,而珊瑚菜素能显著抑制TNF-α激活的黏附分子的表达。该研究结果表明,珊瑚菜素通过抑制HUVECs表面黏附分子表达阻止MCF-7细胞在血管内皮细胞上的黏附。
3.4 珊瑚菜素抑制EMT相关调控因子的转录和表达
EMT转化能够促进肿瘤细胞侵袭和迁移能力,是形成转移的又一关键过程。本研究采用qRT-PCR和Western blot实验检测珊瑚菜素对MCF-7细胞中EMT相关调控因子表达的影响。结果见图4,珊瑚菜素显著抑制EMT通路相关调控因子CDH2、FH1、TWIST、ZEB1、MMP2、SNAIL1、SNAIL2、HIF和ZEB2的mRNA转录水平。此外,珊瑚菜素还能够显著下调TRIM22、THBS1和KITLG的mRNA转录和蛋白表达水平(图5)。该研究结果表明,珊瑚菜素能够通过下调EMT通路相关调控因子的表达,从而抑制肿瘤细胞的EMT转化和转移。
3.5 珊瑚菜素抑制小鼠乳腺癌细胞肺转移
通过BALB/c小鼠乳腺癌肺转移模型评估珊瑚菜素预防乳腺癌转移的作用,结果见图6,珊瑚菜素治疗后小鼠肺转移结节数明显减少且呈药物浓度依赖抑制;与正常组相比,肿瘤组的肺组织质量增加,而经珊瑚菜素治疗后趋于正常。HE染色结果同样表明(图7~8),相较于肿瘤组,经过珊瑚菜素治疗后肺组织中肿瘤结节明显减小或消失,尤其是高剂量实验组的情况已基本恢复正常。此外,通过心、肝、脾、肺、肾HE染色结果可以发现,珊瑚菜素没有产生明显的毒副作用。说明了珊瑚菜素在安全剂量下能够抑制小鼠乳腺癌肺转移。
3.6 珊瑚菜素增强小鼠抗肿瘤免疫活性
为阐明珊瑚菜素是否具有抗肿瘤免疫活性,本研究采用流式细胞仪检测各组小鼠外周血中抗肿瘤免疫细胞细胞毒性T淋巴细胞(cytotoxic T lymphocytes,Tc)和自然杀伤细胞(natural killer cells,NK)细胞比例的变化。结果表明,与正常组相比,肿瘤组小鼠外周血中Tc(图9)和NK(图10)细胞比例明显降低,而给药治疗组小鼠外周血中Tc和NK细胞百分比都高于肿瘤组且呈浓度依赖上升。这说明珊瑚菜素可以提高小鼠外周血中Tc和NK细胞的百分比,增强小鼠抗肿瘤免疫细胞活性。
4 讨论
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乳腺癌的全球发病率逐年上升,是导致女性死亡率最高的恶性肿瘤之一[17],乳腺癌的转移亦是其复发的主要原因。肿瘤转移过程复杂,其中CTCs在血液微环境中存活并逃避免疫细胞攻击、黏附于血管内膜并跨内皮细胞迁移是最终形成转移的最为关键一环,也是最为脆弱的一环[3]。因此,抑制CTCs在血管内皮细胞的黏附和跨内皮细胞迁移、增强抗肿瘤免疫细胞活性或许能够有效抑制肿瘤转移过程[8]。由于抗肿瘤药物通常具有较高细胞毒性,在杀死肿瘤细胞的同时必然会直接抑制肿瘤细胞在血管内皮细胞的黏附。因此,评估肿瘤转移预防药物的疗效方法不同于抗肿瘤药物,需要在几乎无细胞毒性的药物浓度下评价肿瘤细胞与内皮细胞之间的黏附。
近年来,基于中药成分的多靶点调控特性,其抗肿瘤药物开发价值备受关注[18]。本研究发现,珊瑚菜素在无显著细胞毒性和小鼠毒副作用的浓度范围内,可通过多机制协同作用抑制肿瘤转移进程。在细胞层面,珊瑚菜素能够显著抑制炎症因子诱导的内皮细胞表面CAMs的表达阻止肿瘤细胞在血管内皮细胞上的黏附、抑制肿瘤细胞EMT相关基因及蛋白的表达水平。T细胞和NK细胞是人体重要的抗肿瘤免疫细胞,在动物转移模型中,珊瑚菜素治疗后能够显著增强小鼠外周血中T细胞和NK细胞含量,说明具有较强的抗肿瘤免疫活性。上述结果表明,珊瑚菜素通过靶向肿瘤微环境中的血管黏附、EMT进程及免疫调节等多重机制,展现出显著的抗肿瘤转移潜力。
TRIM 22是TRIM家族蛋白的成员之一,可作为E3泛素连接酶可以通过激活NF-κB或MAPK信号传导和加速IκBα或Raf-1降解来促进细胞瘤细胞的生长和转移[19-20]。THBS1在细胞黏附/迁移、血管功能、炎症及肿瘤转移过程中具有重要作用[21-22]。KITLG在肿瘤细胞骨架和迁移调控中发挥重要作用[23-24]。本研究发现珊瑚菜素在无细胞毒性浓度下能够抑制TRIM22、THBS1和KITLG的表达,为进一步研究珊瑚菜素预防肿瘤转移的药理机制提供了新思路。
综上所述,本研究初步阐明了珊瑚菜素在无细胞毒性浓度下能够通过抑制黏附分子的表达阻止CTCs在血管内皮细胞的黏附、抑制EMT相关调控因子的表达、增强小鼠抗肿瘤免疫活性,从而表现出预防肿瘤转移的作用。但珊瑚菜素抑制内皮细胞表面黏附分子表达、抑制肿瘤细胞EMT和增强小鼠抗肿瘤活性的分子机制还未被阐明,需要进一步考察。
基金
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  • 国家自然科学基金项目资助(81703555)
  • 福建省自然科学基金项目资助(2020J01859)
文献
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2025年第60卷第8期
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doi: 10.11669/cpj.2025.08.006
  • 接收时间：2025-01-08
  • 首发时间：2025-11-12
  • 出版时间：2025-04-15
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  • 收稿日期：2025-01-08
基金
国家自然科学基金项目资助(81703555)
福建省自然科学基金项目资助(2020J01859)
作者信息
    1 福建中医药大学药学院, 福建省中药学重点实验室, 福州 350122
    2 福州海洋研究院, 闽江学院材料与化学工程学院, 福州 350108

通讯作者:

*黄鸣清,男,博士,教授,博士生导师 研究方向:中药学 Tel:(0591)22861283;
卢余盛,男,博士,副研究员,硕士生导师 研究方向:抗肿瘤药物药 Tel:(0591)83761795
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https://castjournals.cast.org.cn/joweb/zgyxzz/CN/10.11669/cpj.2025.08.006
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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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