| Background and objective: Uterine cervix cancer is one of the most common malignant tumor in the world,which seriously endangers women’s health.Drug resistance,invasion,metastasis and recurrence in cervical cancer patients are the main cause of women’s death,which also affect the clinical treatment and prognosis.Although advanced cervical cancer patients can not be completely cured,but there are many options can improve the patient’s life length and quality.In recent years,with the rapid development of precision medicine in cancer treatment,the tumor biology can provide a direction for finding new ways to treat cancer.CLCA2 has been identified as one of the target genes of p53 gene,which plays an important role in regulating cell proliferation,migration and invasion.It has been found that CLCA2 is closely related to the invasion and metastasis of various tumors,but its role in cervical cancer is not clear.Our previous study have found that the CLCA2 gene was significantly up-regulated after the treatment of Isocorydine by using gene chip technology.Therefore,the aim of this study was to investigate the effect of CLCA2 Gene in cervical cancer and its possible mechanism,and to explore the sensitivity of CLCA2 to paclitaxel.We hope to provide a solid experimental foundation for the diagnosis and treatment of cervical cancer.Methods: We constructed the specific CLCA2-sh RNA and transferred it into the Siha cells of the cervical cancer by means of lentiviral vector.The experiment was divided into Siha group(blank control group),Siha-NC group(negative control group),CLCA2-sh RNA-1(CLCA2-sh RNA-1transfected cervical cancer Siha cells,hereinafter referred to as K1 group)and CLCA2-sh RNA-2 group(CLCA2-sh RNA-2 transfected cervical cancer Siha cells,hereinafter referred to as K2).The knockout efficiency of CLCA2 gene was confirmed by western blot.The invasion of Siha cells was observed by transwell invasion assay.The expression of E-cadherin,Vimentin,MMP-2 and MMP-9 were detected by immunohistochemistry.Finally,the sensitivity of knockout CLCA2 gene in cervical cancer Siha cells to paclitaxel was investigated by MTS assay.Result: 1.We were successful construction of lentiviral vector,which can be efficiently transferred into cervical cancer Siha cells.Western blot analysis showed that the expression level of CLCA2 protein in K1 group and K2 group was significantly lower than that of Siha group and Siha-NC group(P <0.05).2.Transwell invasion assay showed that the number of invaded cells in K1 group and K2 group was significantly increased compared with Siha group and Siha-NC group,the difference was statistically significant(P <0.05).3.The expression of E-cadherin in K1 and K2 groups was significantly lower than that of Siha group and Siha-NC group(p <0.05),while the expression of vimentin protein in K1 group was significantly higher than that of Siha group(P <0.05),but there was no significant difference compared with Siha-NC group(P> 0.05),K2 group was significantly higher than that of Siha group and Siha-NC group(P <0.05);The further expression of MMP-2 and MMP-9 in K1 and K2 groups was significantly higher than that of Siha group and Siha-NC group(P <0.05).The results of MTS showed that the inhibition rate of K1 and K2 groups in different concentrations of paclitaxel was significantly increased compared with Siha group and Siha-NC group,the difference was statistically significant(P <0.05).Conclusion: 1.CLCA2 gene knockout can enhance the invasive ability of cervical cancer Siha cells;2.CLCA2 gene knockout can promote the epithelial-mesenchymal transition of cervical cancer Siha cells;3.The enhancement of the invasive ability of cervical cancer Siha cells after CLCA2 gene knockout may be achieved by inducing the expression of MMP-2 and MMP-9;4.CLCA2 gene knockout can enhance the sensitivity of paclitaxel to cervical cancer Siha cells. |
PDF Full Text Request