Effect of hepatitis B x gene on the expression of major histocompatibility complex class I chain-related gene A-A5. 1,invasion,and migration of HepG2. 2. 15 cells

Abstract

Objective To investigate the effect of hepatitis B x( HBx) gene on the expression of major histocompatibility complex class I chain-related gene A( MICA)-A5. 1,invasion,and migration of HepG2. 2. 15 cells. Methods HepG2. 2. 15 cells( HepG2 cells with the insertion and continuous expression of whole HBV genome) were cultured in vitro and were randomly divided into control group,HBx overexpression plasmid group,HBx empty plasmid group,HBx siRNA group,and HBx siRNA negative control group. After the transfection of plasmid or siRNA,CCK-8 assay was used to measure cell proliferation after 24 and 48 hours to screen out the appropriate duration of drug action; the Transwell invasion test and wound-healing assay were used to observe the changes in cell invasion and migration abilities;immunoblotting was used to measure the expression of HBx,MICA-A5. 1,and the marker proteins for epithelial-mesenchymal transition E-cadherin and vimentin; ELISA was used to measure the level of soluble MICA( s MICA) in cell culture medium. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK q test was used for further comparison between two groups. Results Compared with the control group after 24 hours,the HBx overexpression plasmid group had a significant increase in cell viability and the HBx siRNA group had a significant reduction in cell viability( q = 8. 268 and 4. 365,P < 0. 001 and 0. 036); compared with the control group after 48 hours,the HBx overexpression plasmid group had a significant increase in cell viability and the HBx siRNA group had a significant reduction in cell viability( q = 12. 680 and 7. 523,both P < 0. 001). Compared with the control group,the HBx overexpression plasmid group had significant increases in the protein expression of HBx,MICA,and vimentin in cells,the level of s MICA in cell culture medium,the number of migrating cells,and the number of cells out of the Transwell chamber,as well as a significant reduction in the expression of E-cadherin( q = 9. 427,6. 697,10. 500,5. 042,22. 740,15. 720,and 5. 258,all P < 0. 05); the HBx siRNA group had significant reductions in the protein expression of HBx,MICA,and vimentin in cells,the level of s MICA in cell culture medium,the number of migrating cells,and the number of cells out of the Transwell chamber,as well as a significant increase in the expression of E-cadherin( q = 8. 133,8. 828,7. 616,7. 673,5. 391,7. 694,and 6. 226,all P < 0. 05). Conclusion The HBx gene regulates theexpression of MICA-A5. 1 in HepG2. 2. 15 cells and the invasion and migration of HepG2. 2. 15 cells,and upregulation of the HBx gene can promote the expression of MICA-A5. 1 and enhance the invasion and migration abilities of HepG2. 2. 15 cells.
- hepatitis B virus,
- Hep G2 cells,
- genes,MHC classⅠ

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Effect of hepatitis B x gene on the expression of major histocompatibility complex class I chain-related gene A-A5. 1,invasion,and migration of HepG2. 2. 15 cells

DOI: 10.3969/j.issn.1001-5256.2020.04.020