The Influence Of Radio-Sensitivity By ADAM9 On Hepatocellular Carcinoma Cell

Objective: In recent years,the efficacy of radiotherapy for hepatocellular carcinoma has become increasingly significant.However,the efficacy of radiotherapy for hepatocellular carcinoma in China needs to be further improved.The main influencing factors are as follows: 1.Hepatocellular carcinoma belongs to a kind of moderately radiosensitive tumor;2.The liver belongs to a radiosensitive organ;in addition,most patients with hepatocellular carcinoma in China are complicated with viral hepatitis,and their basis of liver functions is poor.The radiation dose received by the liver may cause more severe radiation-induced liver injuries before reaching the therapeutic goal of damaging hepatocellular carcinoma cells.Therefore,elucidating the relevant mechanisms regulating the radio-sensitivity of HCC cells,increasing the radio-sensitivity of HCC cells,and improving the efficacy of radiotherapy for HCC are still hot issues to be solved urgently.ADAM9 is a zinc-dependent transmembrane protein that widely exists on the cell surface.Several studies have shown that ADAM9 can promote the development of liver cancer,but its relationship with radio-sensitivity of liver cancer cells has not been reported.In this experiment,ADAM9 protein of hepatoma cells was reduced or overexpressed by lentiviral infection,and the changes of cell biological behaviors such as cell proliferation,subcutaneous tumorigenesis in nude mice and the expression of related proteins were observed after X-ray irradiation.The relationship between ADAM9 and radio-sensitivity of hepatoma cells was explored to provide theoretical support for finding new molecular targets for enhancing radio-sensitivity of hepatoma cells.Methods: To construct lentiviral vector of ADAM9 gene overexpression and knockout,ADAM9 knockout lentivirus vector was transfected into the hepatoma cell line MHCC97 H and compared with the control group.In the same way,ADAM9 over expressed lentivirus vector was transfected into Huh-7 cell line,and compared with the control group.After 4Gy X-ray irradiation,CCK-8 was used to detect the cell proliferation.The colony forming ability was studied by agar clone forming experiment,and the growth ability was detected by subcutaneous tumor forming experiment in nude mice.Thereafter,statistical analysis was carried out.Western blot was used to detect the expression of apoptosis-related proteins Bcl-2,Bax and DNA damage-related proteinγ-H2 AX,and statistical analysis was performed.Result: 1.After ADAM9 was knocked out,the expression of ADAM9 protein in MHCC97 H liver cancer cells was significantly inhibited;after overexpression of ADAM9,the expression of ADAM9 protein in liver cancer Huh-7 cells was enhanced.2.The result of CCK-8 showed that by comparing with the idling group,the cell proliferation of the knockout group of ADAM9 protein was significantly lower(P < 0.01),such difference was statistically significant;by comparing with the idling group,the cell proliferation of the liver cancer cell Huh7 overexpression group was significantly increased(P < 0.01),such difference was statistically significant.3.It was found by agar clone formation experiments that by comparing with the idling group,the ADAM9 protein knockout group had less ability to form clones(P<0.01);by comparing with the idling group,the overexpression group had stronger clone formation ability(P<0.01);4.It was found by the subcutaneous tumor formation experiments in nude mice that by comparing with the idling group,the ADAM9 protein knockout group had an ability of reducing subcutaneous tumor formation ability,and such difference was statistically significant(P<0.01);by comparing with the idling group,the overexpression group had a better subcutaneous tumor formation ability(P<0.01).5.Western blot results showed that compared with air subletting,the expression of Bcl-2 protein was significantly reduced after knocking out ADAM9 protein,and the expression of Bax and γ-H2 AX proteins was significantly increased;compared with the control group,Bcl-2 protein Decreased expression and increased Bax and γ-H2 AX protein expression.Therefore,Knockdown of ADAM9 protein can significantly inhibit cell proliferation,colony formation and tumorigenicity of hepatocellular carcinoma cells after radiotherapy;overexpression of ADAM9 protein can enhance growth ability of hepatocyte cells after radiotherapy.Conclusion: Knockdown of ADAM9 protein can significantly inhibit cell proliferation,colony formation and tumorigenicity of hepatocellular carcinoma cells after radiotherapy;overexpression of ADAM9 protein can enhance growth ability of hepatocyte cells after radiotherapy.Therefore,ADAM9 protein can inhibit the radio-sensitivity of liver cancer cells.