The Effect Of CLIC1 On The Expression Of E-Cadherin And Vimentin In Cervical Cancer SiHa Cells And Its Relationship With EMT

In the global list of gynecological malignancies,cervical cancer stands the first on the list and the newincidence increases year by year.The age of onset tends to be younger than before.With the highest incidence in China,cervical cancer seriously affects women's quality of life.Cervical cancer can be further developed from in situ to invasive or metastatic carcinoma,which is an important factor leading to reduced survival rate and even death of patients.Therefore,to explore the pathogenesis and progression mechanism of cervical cancer is very important to the diagnosis,treatment and prognosis of cervical cancer,especially for improving the survival rate and prognosis of patients.CLIC1,also known as chloride nclear channel-27(NCC27),has a relative molecular of about 27,000 and chloride ion channel function.Studies have confirmed that CLIC1 is highly expressed in a variety of malignant tumor tissues,such as gastric cancer,colon cancer,liver cancer,gallbladder cancer etc,and its high expression is closely related to tumor cell migration and invasion,lymphatic metastasis,malignant transformation and tumor stem cell resistance,suggesting that CLIC1 may be an important factor in prognosis.Studies have shown that epithelial-mesenchymal transformation can promote the invasion and metastasis of cervical cancer cells,and promote malignant transformation of cervical lesions.The overexpression of CLIC1 can reduce the expression of e-cadherin and increase the expression of Vimentin in the epithelial-mesenchymal transformation,which is consistent with the occurrence of epithelial-mesenchymal transformation.In our study,CLIC1 overexpressed recombinant plasmid was transfected into cervical cancer Si Ha cells and si RNA interference technology was applied to study the occurrence and development mechanism of cervical cancer at the cellular level,so as to provide certain basis for the early diagnosis andtreatment of cervical cancer.Objective:To investigate the effect of changing the expression of CLIC1 on the expression of E-cadherin and Vimentin in cervical cancer Si Ha cells,and further analyze the relationship between the expression of CLIC1 and EMT of cervical cancer Si Ha cells.Methods:1.Resuscitation and subculture of cervical cancer Si Ha cells;2.Transfection of CLIC1 overexpression recombinant plasmid and CLIC1 si RNA;3.The expression level of CLIC1,E-cadherin,and Vimentin m RNA of the cells(Transfection overexpression plasmid group;Transfection blank plasmid group;Untreated blank control group;si RNA transfection group;si RNA negative control group)were detected by RT-q PCR.4.The protein expression levels of CLIC1 protein,E-cadherin and Vimentin of the five group were detected by Western blot;5.All data was analyzed by SPSS19.0 statistical analysis software,SNK method and LSD method were used for statistical analysis between groups and within groups.Results:1.RT-q PCR results showed that the relative expression level of CLIC1 m RNA in the overexpressed plasmid group(1.97 ± 0.08)was higher than that in the blank plasmid group(1.15 ± 0.02)and the blank control group(1.10 ± 0.01),and the relative expression level of CLIC1 m RNA in the Si RNA transfection group(0.67 ± 0.03)was lower than that in the blank control group(1.10 ± 0.01)and the si RNA negative control group(1.17 ±0.02),the difference was statistically significant(P<0.01).2.The relative expression of Vimentin m RNA in overexpressed plasmid group(0.98 ±0.04)was higher than that in blank plasmid group(0.58 ± 0.04)and blank control group(0.53 ± 0.06),and the relative expression of E-cadherin m RNA(0.32 ± 0.04)was lower than that in the blank control group(0.62 ± 0.04)and the si RNA negative control group(0.58 ± 0.05),and the difference was statistically significant(P<0.01).3.The relative expression of E-cadherin m RNA in Si RNA transfection group(1.12 ± 0.06)was higher than that in the blank control group(0.58 ± 0.05)and the si RNA negative control group(0.64 ± 0.04),and the relative expression of Vimentin m RNA(0.39 ± 0.05)was lower than that of the blank control group(0.53 ± 0.06)and the si RNA negative control group(0.54 ± 0.04),and the difference was statistically significant(P<0.01).However,there were no statistical significance of the relative expression levels of CLIC1,e-cadherin and Vimentin m RNA among the blank plasmid group,blank control group and si RNA negative control group(P > 0.05).4.The expression level of CLIC1 protein in the overexpression plasmid group(1.48 ±0.06)was higher than that in the blank plasmid group(0.71±0.12)and the blank control group(0.66±0.07);the protein expression level of vimentin in the overexpression plasmid group(0.69 ± 0.06)above the blank plasmid group(0.40 ± 0.10)and the blank control group(0.39 ± 0.09),while the expression level of e-cadherin protein(0.23 ± 0.07)was lower than that in the blank plasmid group(0.36 ± 0.06)and the blank control group(0.34 ± 0.04),the difference was both statistically significant(P<0.01).5.The expression level of CLIC1 protein in Si RNA transfection group(0.45± 0.02)was lower than that in blank control group(0.80 ± 0.03)and si RNA negative control group(0.85 ± 0.04),and the relative expression of Vimentin(0.26 ± 0.04)was lower than that of the blank control group(0.39 ± 0.09)and the si RNA negative control group(0.40 ±0.05),the relative expression of e-cadherin(0.89 ± 0.07)was higher than that of the blank control group(0.34 ± 0.04)and the si RNA negative control group(0.35 ± 0.07),and the difference was statistically significant(P<0.01).However,there were no statistical significance of the relative expression levels of CLIC1 protein,e-cadherin and Vimentinamong the blank plasmid group,blank control group and si RNA negative control group(P > 0.05).Conclusion:1.CLIC1 overexpression in cervical cancer Si Ha cells can promote the occurrence of EMT in cervical cancer Si Ha cells.2.Blocking the expression of CLIC1 may have positive clinical significance in the treatment of cervical cancer.