Synergistic Inhibitory Effect Of RMP And ELP3 On ⁶⁰Coγ-ray-induced Apoptosis Of Hepatocellular Carcinoma Cells

Objective:To study the effect of RMP and ELP3 on the apoptosis of hepatocellular carcinoma cells induced by irradiation and to explore the molecular mechanism of RMP and ELP3 in this process.Methods:1.Construction of RMP overexpression and interference plasmid.2.The effect of RMP or ELP3 on the expression of apoptosis-related proteins was detected by immunohistochemistry in nude mice subcutaneously inoculated with HCC cells.3.The effect of ELP3 on the expression of apoptosis-related proteins induced by irradiation was detected by immunohistochemistry.4.Immunohistochemistry was used to detect the expression of RMP and ELP3 in hepatocellular carcinoma and paraneoplastic tissues.5.The correlation between RMP and ELP3 protein expression in hepatocellular carcinoma cell lines was detected in vivo and in vitro.6.CCK-8 was used to detect the effect of RMP and ELP3 on the proliferation of HCC cells.7.The effect of RMP and ELP3 on DNA damage repair of irradiated hepatocarcinoma cells induced by comet assay.8.The effect of RMP and ELP3 on the apoptosis of hepatocellular carcinoma cells induced by irradiation was detected by flow cytometry.9.Immunoprecipitation was used to detect the interaction between RMP and ELP3.Results:1.Successfully construct RMP overexpression and interference plasmid.2.In vivo,RMP and ELP3 could promote the expression of apoptotic gene in hepatocellular carcinoma cells.3.In vivo,ELP3 promotes the expression of apoptosis-inducible protein in irradiated hepatocellular carcinoma cells.4.In pathological tissues,RMP or ELP3 is highly expressed in HCC tissues.5.There was no correlation between RMP and ELP3 in protein expression levels in vivo and in vitro.6.RMP or ELP3 can promote the proliferation of hepatocarcinoma cells,the two co-expression effect after the superposition effect.When one side is over-expressed and the other is interfered,the gain effect of overexpression will be significantly inhibited.7.RMP or ELP3 could promote DNA damage repair of hepatocellular carcinoma cells,and the effect of RMP or ELP3 was superimposed.When one side is over-expressed and the other is interfered,the gain effect of overexpression will be significantly inhibited.8.RMP or ELP3 could inhibit the apoptosis of hepatocarcinoma cells induced by irradiation,and the effect of RMP or ELP3 was superimposed.When one side is over-expressed and the other is interfered,the gain effect of overexpression will be significantly inhibited.9.RMP and ELP3 are present in the form of complexes in hepatocellular carcinoma cells.The level of binding of RMP to ELP3 increases after irradiation.Conclusion:RMP and ELP3 promote the proliferation of hepatocellular carcinoma cells in the form of complexes,promote radiation-induced DNA damage repair and inhibit subsequent apoptosis.