Molecular Mechanism Of Inhibitory Effect Of GINS2-shRNA On Malignant Melanoma

Objective: To observe the effect of GINS2-shRNA(shGINS2)on malignant melanoma xenograft model in nude mice,and to explore the molecular mechanism of GINS2 inhibition on malignant melanoma.Methods:1.Cell experiment: Cultured A375 cells transfected with shGINS2 and shNC;Detected the expression of GINS2,P-P38 and caspase-3 proteins in A375 cells transfected with shGINS2 and shNC by Western Blot.2.Animal experiment: A375 cells transfected with shGING2 and shNC were inoculated into the unilateral armpit of nude mice to prepare a malignant melanoma xenograft model in nude mice,and divided into experimental group: shGING2 group and negative control group;The growth of the tumor was observed daily,and the size of the transplanted tumor was recorded when the tumor emerged,and the tumor growth curve was drawn;After continuous observation for 3 to 4 weeks,the nude mice were sacrificed to remove the complete tumor tissue,the tumor volume and tumor weight were measured,and the tumor inhibition rate was calculated;Detected the expression of GINS2,P-P38 and caspase-3 proteins in tumor tissues by Western Blot.3.Statistical analysis the results of each group.Results: 1.Cell experiment: Western blot results showed that after shGINS2 was transfected into A375,the expression level of GINS2 protein in shGINS2 group was significantly lower than that in shNC group,while the protein expression levels of P-P38 and caspase-3 were significantly increased(P<0.001);2.Animal experiments: none of the experimental animals died.From the tumor growth curve,it can be seen that compared with the shNC group,the shGINS2 group had an inhibitory effect on the growth of the transplanted tumor.At the end of the intervention,the tumor volume and tumor weight in the shGINS2 group were significantly reduced compared with those in the shNC group(P<0.001);the shGINS2 group had a higher tumor inhibition rate,with a tumor volume inhibition rate of 61.33% and a tumor weight inhibition rate of 61.17%;Western blot results showed that in tumor tissue,the expression of GINS2 protein in the shGINS2 group was lower than that in the shNC group,while the expressions of P-P38 and caspase-3 proteins were increased(P<0.001).Conclusion: The results of this study confirmed that shGINS2 has inhibitory effect on the xenograft model of malignant melanoma in nude mice;the inhibitory mechanism of shGINS2 on malignant melanoma may be by activating the p38 MAPK signaling pathway,thereby initiating the p38MAPK/caspase-3 cascade reaction,and then promoting the achieved by apoptosis.