Effect And Mechanism Of C3G Gene On Apoptosis And Proliferation In H9C2 Cardiomyocytes

Objectives:To study the effect and mechanism of C3G gene on apoptosis and proliferation in H9C2 cardiomyocytes.Methods:Design RNA interference sequence, purchase pCXN2-Flag plasmid (empty plasmid) and pCXN2-Flag-hC3G plasmid (overexpression of hunman C3G mRNA). Then H9C2 cardiomyocytes were randomly infected and transfected with blank reagent、negative lentivirus、C3G siRNA lentivirus、C3G siRNA lentivirus+pCXN2-Flag plasmid and C3G siRNA lentivirus+pCXN2-Flag-hC3G plasmid. So the experiment were randomly divided into five groups, namely blank group、negative control group、C3G silence group、C3G silence+empty plasmid group and C3G silence+C3G overexpression group. After 72 hours from transfection plasmid, the expression of C3G mRNA was detected by PT-RCR,cell apoptosis was analyzed by flow cytometry, cell proliferation rate was determined by MTT, using Western blotting tested the protein level of C3G、p-ERK1/2、Bax and Flag.Results:Fluorescence graph:lentivirus infection by puro filter more than 85% of the cells are marked by green fluorescent protein, namely more than 85% of the cells were infected lentivirus. Compared with the blank group and negative control group, the Bax protein and cell apoptosis rate were significantly increased in C3G silence group and C3G silence+empty plasmid group(P<0.01),though its C3G mRNA、C3G protein、p-ERK1/2 protein and cell proliferation rate were remarkable decreased(P<0.01, P<0.05); On the contrary, C3G silence+C3G overexpression group has the opposite results,respectively.Conclusions:C3G gene silence can induce apoptosis and inhibit proliferation in H9C2 cardiomyocytes, overexpression of C3G gene to reverse C3G silence influence in H9C2 cardiac muscle cells, characterized by reduced apoptosis and promoted proliferation; which may be related to p-ERK1/2 protein and pro-apoptosis molecule Bax.