RACK1 Affect The Ability To Attack Of Cervical Cancer Cells SiHa In Vitro Through RhoA Kinase

Objective:1.To detect the expression levels of RACK1 and RhoA in the overexpression plasmid group,the blank plasmid group,and the untreated group of cervical cancer cell strains SiHa,and to explore the correlation between them;2.To detect the expression of RhoA in the overexpression plasmid group,the blank plasmid group and the untreated group SiHa after treatment with different concentration gradients of RhoA kinase inhibitor,and the invasiveness was measured.Methods:1.Cultivate SiHa cells,lipid-mediated,a set of transfected over-expression plasmids,one set of transfected blank plasmids,and the other set without any treatment,Real-time qPCR and Two Western blot methods were used to detect the expression of RACK1 and RhoA in the three groups of SiHa;2.The expression of RhoA was detected in three groups after treatment with different concentration gradients of RhoA kinase inhibitors.The invading ability of the three groups was detected again by transwell chamber;3.Analysis using SPSS 16.0 software,all data are expressed as mean ±standard deviation(x ± s)form,P <0.05 for statistical significance.Results:1.The expression levels of RACK1 and RhoA in the overexpression plasmid group detected by PCR were 1.25±0.59 and 1.36±0.04,The expression levels of RACK1 and RhoA in the overexpression plasmid group detected by western blot were 0.94±0.08 and0.52 ± 0.07,which was higher than the other two groups,There was no significantdifference in the expression levels of RACK1 and RhoA between the blank plasmid group and the untreated group.Pearson correlation coefficient analysis that RACK1 and RhoA are both positively correlated,and the correlation coefficient r is 0.929,0.601;2.Y-27632 with different concentration gradients inhibited RhoA kinase in the three groups,with the increase of inhibitor concentration,the expression and invasive ability of RhoA decreased.Conclusions:1.Successfully transfected overexpression plasmids in cervical cancer SiHa cells,and simultaneously obtained SiHa cells with stable overexpressed RACK1;2.Overexpression of RACK1 affect the invasion ability of cervical cancer cell line SiHa by RhoA kinase.