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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.
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| 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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