Objective: To determine the influence of HBx gene RNA interference combined with chemotherapy on stable hepatocellular carcinoma cells growth and its apoptosis mechanism.
Methods: Stable hepatocellular carcinoma cells transfected by shRNA aiming at HBx together with independent control series (MHCC97-H,HK3, and 21543) were identified. The extent of HBx gene by RNA interference was detected by RT-PCR. The influence of cell growth through RNA interference was observed with cell counting kit-8 (CCK8), the diversity of cell cycle by flow cytometry and cell apoptosis were detected by TUNEL apoptosis detection kit.
Results: RT-PCR demonstrated that the HBx mRNA level of 21,543 cell down regulation was 91%. The HBx mRNA level of HK3 cells was not different from MHCC97-H cell. The growth of 21,543 cells was obviously slower than MHCC97-H cells and HK3 cells, with no significant difference. The cell cycle of 21,543 cells showed that hepatocellular carcinoma cells through RNA interference targeting at HBx delayed in go to S stage, and the proliferation activity degraded obviously. The 3 kinds of cells adding different concentrations of flurouracil and cisplatin grew slowlier than the origin cells. The growth inhibition was dependent on the concentration of drug with growth inhibition of 21,543 cells the most obvious.That of the 3 kinds of cells adding alpha-interferon was not obvious.Flurouracil induced apoptosis in all cells. Apoptosis in 21,543 cells was the most obvious.
Conclusion: RNA interference targeting at HBx can suppress the growth of hepatocellular carcinoma cells. Hepatocellular carcinoma cells through RNA interference targeting at HBx can intensify chemo-sensitivity. Combination of RNA interference targeting at HBx with chemotherapeutics can induce apoptosis in more hepatocellular carcinoma cells and cell proliferation steps down accordingly.