Experiment on Inhibiting NEK7 to Promote Apoptosis of Hepatocellular Carcinoma Cells

Abstract

Objective To use in vitro experiments to verify the changes of proliferation, senescence and apoptosis of hepatocellular carcinoma cells after inhibiting the expression of NEK7, and to explore the related molecular mechanism. Methods Western blot and RT-PCR were used to detect the expression of NEK7 in hepatocellular carcinoma cells and THLE-2 cells. A viral vector was designed to inhibit the expression of NEK7 based on the gene sequence. After hepatocellular carcinoma cells were transfected, we observed the changes of proliferation activity, cell senescence, cell apoptosis and cell cycle in vitro. Western blot was used to detect the expression of cell cycle-related factors. Results Compared with THLE-2 cells, NEK7 was highly expressed in hepatocellular carcinoma cells. After inhibiting the expression of NEK7 with shRNA, the proliferation of hepatocellular carcinoma cells was inhibited, the proportions of cell senescence and apoptosis were increased, meanwhile, the cell number in stage S and G2/M was significantly reduced, the cell cycle progression was blocked, the expression levels of C-myc, c-Fos, cyclin D1 and cyclin E were inhibited, P16 and P27 expression were increased, and CDK2, CDK4 and CDK6 expression were not significantly changed. Conclusion After inhibiting the expression of NEK7, the proliferation ability of hepatocellular carcinoma cells is reduced, cell senescence is promoted and apoptosis is induced; meanwhile, the cell cycle progress is blocked.

Keywords

• hepatocellular carcinoma
• nek7
• cell cycle