Effect Of RACK1 On The Proliferation And Apoptosis Of SiHa Cell

Objective:Activated protein kinase receptor 1(RACK1)is a dissociative scaffolding protein in cells,which participates in many biochemical reactions and affects the biological functions of cells.Previous studies have confirmed that in cervical cancer,the expression of RACK1 was significantly higher than that in normal tissue,and positive correlation was found in normal tissues,cervical intraepithelial neoplasia I,II,III and cervical cancer tissues.Based on previous research results,this experiment mainly verify that the effects of increase of RACK1 expression on cervical cancer SiHa cell proliferation,apoptosis ability,and lay the foundation for the further study of RACK1 in cervical cancer development and prognosis of disease mechanisms.Methods:The human SiHa cell is provided by Shanghai cell bank,and it is selected for subculture,and transfected by RACK1 overexpressing plasmids and blank plasmids.After transfection,The relative expression of mRNA of RACK1 protein in all cells after transfection is measured by RT-qPCR,and the relative expression of RACK1 protein in each cell is measured by Western-Blot method,and a stable high expression of RACK1 cell is obtained.Then CCK-8 method is used to continuously measure the absorbance of the cells after transfection by the enzyme labeling instrument,in order to compare the proliferation of each group.Finally,cell apoptosis is detected by flow cytometry.At the last,using statistical data of the SPSS16.0 software analysis data.Results:1.The relative expression of mRNA measured by RACK1 overexpressed plasmid group is 1.25±0.59,compared with the blank plasmid transfection group(0.71±0.12)and the untransfected group(0.66±0.07),the content increased significantly(P< 0.05).2.The relative content of RACK1 protein in the overexpressed plasmid group(0.94 ±0.08)is higher than the other groups,and the difference between the two groups is statistically significant(P < 0.05),but there is no significant difference between the blank plasmid group(0.74 ± 0.07)and the relative content of RACK1 in the untreated group(0.72± 0.09).3.Overexpression of RACK1 can promote the proliferation of SiHa cell in cervical cancer.Continuous measurement of cell absorbency in each group by CCK-8 method,then we can compare the absorbance between each group.And we can find the cell proliferation of the expression plasmid group is obviously higher than that of the other two groups(P < 0.05).4.The overexpression of RACK1 can inhibit the apoptosis of cervical cancer cells.The apoptosis rate(5.70 ± 1.42)is lower than that of the blank plasmid transfection group and the untransfected group(15.73 ± 1.29,17.37±3.01(P < 0.05)).Conclusion:1.Successfully constructed RACK1 overexpressing cervical cancer cell line SiHa in vitro;2.The over expression of RACK1 can promote the proliferation of cervical cancer cells in vitro,and inhibit cell apoptosis,Suggesting that it plays a certain role in advancing the progress of cervical cancer.