Effect Of MiR-142-3p On Apoptosis Of Liver Cancer Cells

Objectivc:Investigate the effect of miR-142-3p on the expression of liver cancer cells in gene expression,the effects of miR-142-3p on the expression of miR-142-3p in liver cancer cells were investigated.Methods:1.Successfully transfected liver cancer cells SK-Hep1 and Hep3B,respectively,the four groups were the overexpressed miR-142-3p group,the inhibited miR-142-3p expression group,the miR-142-3p control group and the normal cells group.2.Overexpressed miR-142-3p group,inhibited miR-142-3p expression group and miR-142-3p control group were detected the apoptosis of SK-Hepl and Hep3B cells by flow cytometiy.3.IRAK1,TRAF6,RIP1,MLKL,caspase8,caspase9,PI3K and BAD proteins were detected the expression of in SK-Hep 1 cells in each group by Western blot and the results were to explore their relationship with apoptosis and necroptosis.Results:1.The percentage of apoptosis in SK-Hepl cells was detected by flow cytometry.The percentage of apoptosis in the overexpressed miR-142-3p negative control group was 8.95%,and the percentage of apoptosis in the overexpressed miR-142-3p group was 20.7%.The difference between the two groups was statistically significant(P<0.01);the percentage of apoptosis in the inhibited miR-142-3p expression negative control group was 28.37%,and the percentage of apoptosis in the inhibited miR-142-3p expression group was 18.68%.The difference was statistically significant between the two groups(P<0.01).2.The percentage of apoptosis in Hep3B cells was detected by flow cytometry.The percentage of apoptosis in the overexpressed miR-142-3p negative control group was 6.91%,and the percentage of apoptosis in the overexpressed miR-142-3p group was 9.77%.The differerice was statistically significant(P<0.05);the percentage of apoptosis in the inhibited miR-142-3p expression negative control group was 31.44%;the percentage of apoptosis in the inhibited miR-142-3p expression groupwas 10.71%.In this comparison,the difference was statistically significant(P<0.01).3.We can detect by western blot that the protein expression levels of IRAK1,TRAF6 and RIP 1 were significantly decreased in SK-Hepl cells after overexpression of miR-142-3p in SK-Hepl cells,and the difference was statistically significant(P<0.01,P<0.05).MLKL,caspase8,caspase9,PI3K,BAD protein expression levels did not change significantly and the difference was not statistically significant(P>0.05).After inhibiting the expression of miR-142-3p,the protein expression levels of IRAK1,TRAF6 and RIP1 were significantly higher than those of the negative control group,the difference was statistically significant(P<0.01,P<0.05),MLKL,caspase8,caspase9.There was no significant change in the protein expression levels of PI3K and BAD,and the difference was not statistically significant(P>0.05).Conclusion:1.miR-142-3p can promote apoptosis and may play a role as a tumor suppressor gene in liver cancer cells.2.In liver cancer cells,miR-142-3p is a negative regulator of apoptosis-related proteins such as IRAK1,TRAF6 and RIP1.3.In liver cancer cells.miR-142-3p has no regulatory effect on apoptosis-related endogenouspathway-related proteins such as PI3K,BAD,caspase9 and necroptosis-related proteins such as MLKL and caspase8.