Effect Of CLIC1 On The Growth,Proliferation And Apoptosis Of Cervical Carcinoma SiHa Cells

Objective:Chloride Intracellular channel 1(CLIC1)is a transmembrane protein.Several studies have reported that CLIC1 is up-regulated in malignant tumors,and some studies have shown that CLIC1 is highly expressed in malignant tumors and is associated with poor prognosis.These evidences suggest that CLIC1 may be an important prognostic factor.The aim of this study is to clearify the effects of CLIC1 on the growth,proliferation and apoptosis of cervical cancer cell line,SiHa,and explore the relationbetween CLIC1 overexpression and and the development or prognosis of cervical cancer.Methods:The human SiHa cell line purchased from Shanghai Cell Bank and was cultured in 10% DMEM.The CLIC1 overexpression plasmid,blank plasmid,CLIC1 siRNA plasmid and siRNA negative control plasmid were transfected by cationic liposome reagent.RT-qPCR and Western-Blot were used to detect the relative expression of CLIC1 mRNA and protein respectivity.In order to compare the proliferation of each group,CCK-8 was used to measure the absorbance of the cells by Microplate Reader.Finally,flow cytometry assay was used to detect cell apoptosis.SPSS 23.0 was used for statistical analyses,and P< 0.05 indicated statistical significance.Results:1.The relative expression of mRNA in CLIC1 overexpressing plasmid group was 1.97 ± 0.08,which was higher than that in blank plasmid transfection group(1.15 ± 0.02)and untransfected group(1.10 ± 0.01),P<0.05.The CLIC1 siRNA group(0.67 ± 0.03)was lower than the siRNA negative control group(1.17 ± 0.02),P < 0.05;2.The relative expression of CLIC 1 in overexpression plasmid group was 1.48 ± 0.06,which was higher than the blank control group(P<0.05),and the CLIC1 siRNA group(0.45±0.02)was lower than the siRNA negative control group(P<0.05).Compared with the blank plasmid group,there was no significant difference among untreated group(0.80 ± 0.03)and the siRNA negative control group(0.85 ± 0.04),P<0.05;3.The absorbance of each group was continuously measured by CCK-8 method.Compared with the blank plasmid group,the proliferation ability of the overexpression group was increased(P<0.05),and the proliferation ability of the siRNA group was lower than that of the siRNA NC group(P<0.05).The cell cycle of each group was determined by cytometry,The number of cells in the G2/M phase increased in the overexpression group(P<0.05),and the number of cells in the G2/M phase decreased in the siRNA group(P<0.05).4.The apoptosis rate of each group after transfection was determined by flow cytometry.The apoptosis rate of over-expressing CLIC1 group was 8.60±1.42,which was reduced comparing to blank plasmid transfection group(116.70±1.42)and untransfected group(17.31±2.55),P < 0.05,compared with the siRNA negative control group(17.46 ± 3.32),the apoptosis rate of the siRNA group increased(25.17 ± 2.51),P < 0.05.Conclusion:CLIC1 protein can promote the proliferation of cervical cancer SiHa cells and inhibit cell apoptosis,suggesting that it can promote the development of cervical cancer.