Effect Of Kiss-1 Gene On Proliferation,Invasion And Migration Of Human Colorectal Cancer Cell Line HCT116 By Rac1 Pathway

[Object] 1.To investigate the effect of Kiss?1 gene on the expression of Rac1 and Rac1?GTP protein in colorectal cancer cells.2.explore whether Kiss?1 can inhibit the proliferation,invasion and migration of colorectal cancer cells through the Rac1 pathway.[Method] 1.The humanized colorectal cancer HCT116 cells were infected by the lentiviral packaging Kiss?1 overexpression vector to obtain stable overexpressing cell lines.The expressions of Kiss?1 mRNA and Kiss?1 protein were detected by RT?PCR and immunoblotting respectively to determine whether Kiss?1 could be stably expressed in each group.2.The levels of Rac1 and Rac1 activation(Rac1?GTP)protein in blank control group,negative control group and Kiss?1 overexpression group were detected by immunoblotting.3.Kiss?1 overexpressing cell lines were treated with Rac1 agonist PDGF.The expression levels of Rac1?GTP in blank control group,negative control group,Kiss?1 overexpression group and Kiss?1 overexpression + Rac1 agonist group were detected by immunoblotting.And CCK?8 method and Transwell chamber were used to detect the proliferation and invasion and migration of the above four groups.[Results] 1.The expression level of Kiss?1 mRNA and Kiss?1 protein was significantly increased after transfection with Kiss?1 overexpressing vector in HCT116 cells,and Kiss?1 was stably overexpressed in cells.2.The results of immunoblotting showed that the expression level of active Rac1 protein in Kiss?1 overexpression group was significantly lower than that in blank control group and negative control group,while the expression level of Rac1 protein was not significantly different between the groups.3.The expression level of Rac1?GTP(Rac1 active protein)was significantly increased after treatment with Rac1 agonist PDGF,The results of CCK?8 showed that the proliferation of Kiss?1 overexpression group was significantly lower than that of blank control group and negative control group at different time points(P<0.05).Transwell chamber invasion and migration experiments showed that the invasion and migration ability of Kiss?1 overexpression decreased significantly(P <0.05).and the proliferation ability of Kiss?1 overexpression group was significantly higher than that of Kiss?1 overexpression group(P <0.05),and the invasion and migration ability of the cells was also enhanced(P <0.05).[Conclusion]Kiss?1 gene inhibits the proliferation,invasion and migration of colorectal cancer HCT116 cells may be related to the Rac1 pathway and may play a role in reducing the activity of Rac1.