To Investigate The Effect Of MiR-30c-5p On The Proliferation And Invasion Of Colon Cancer Cell By Regulating EDNRA Gene

Objective:To observe the biological behavior of miR-30c-5p in inhibiting the proliferation and invasion of colon cancer cells by targeting EDNRA gene and to explore its mechanism,so as to provide a basis for further study of the function of miR-30c-5p in colon cancer.Method:1.Tumor tissues and adjacent tissues of 20 patients with colon cancer were collected.The expression of miR-30c-5p in colon cancer tissues and adjacent tissues was detected by qRT-PCR,and the correlation between miR-30c-5p expression level and clinicopathological parameters was analyzed.2.qRT-PCR was used to detect the expression of miR-30c-5p in normal colon cell line(FHC)and colon cancer cell lines(SW480,SW620,HT-29,HT-116,LOVO).miR-30c-5p inhibitor was transfected into colon cancer cell lines by liposome transfection,and the transfection efficiency was verified by qRT-PCR.CCK-8 and Transwell were used to detect the effect of down-regulation of miR-30c-5p on the proliferation and invasion of tumor cells.3.mirTarbase website predicted that miR-30c-5p had a targeted binding relationship with EDNRA.GEPIA2 and ENCORI websites were used to verify the expression of EDNRA in colon cancer tissues and its correlation with miR-30c-5p.qRT-PCR and Western blot were used to verify the expression of EDNRA in colon cancer cell lines at mrna and protein levels.Dual luciferase assay was used to verify the target binding sites of the two.4.CCK-8,Transwell and Western blot assays were used to detect the changes in the proliferation,invasion and EMT related indicators of colorectal cancer cells co-transfected with miR-30c-5p inhibitor and EDNRA inhibitor.Results:1.Compared with adjacent tissues,the expression of miR-30c-5p was down-regulated in colon cancer tissues,and the low expression of miR-30c-5p was related to lymph node metastasis,distant metastasis,positive vascular invasion,and TNM stage(Ⅲ,Ⅳ)(all P <0.05).2.The expression level of miR-30c-5p was decreased in colon cancer cells.Compared with the NC group,the cell proliferation and invasion ability were significantly enhanced in the Mir-30c-5P-inhibitor group(P<0.05).3.EDNRA was highly expressed in colon cancer tissues and colon cancer cells,and was negatively correlated with miR-30c-5p(P<0.05).Compared with the NC group, qRT-PCR and WB experiments verified that the expression of EDNRA was up-regulated in the miR-30c-5p-inhibitor group(P<0.05).Dual luciferase assay was used to further verify the targeting relationship between them.4.Compared with the NC group,the cell proliferation and invasion ability of the siEDNRA group were significantly decreased,and the cell proliferation and invasion ability of the miR-30c-5p-inhibitor+siEDNRA group were significantly increased(P<0.05).Compared with the control group,the expression of E-cadherin in the siEDNRA group was increased,and the expression of N-cadherin,Vimentin and EDNRA was decreased.Compared with the siEDNRA group,the expression of E-cadherin was decreased,and the expressions of N-cadherin,Vimentin and EDNRA were increased in the miR-30c-5p-inhibitor+siEDNRA group(all P <0.05).Conclusion:1.The expression of miR-30c-5p is low in colon cancer tissues,which is negatively correlated with the malignant clinicopathological characteristics of colon cancer,and plays a tumor suppressor role in colon cancer.2.miR-30c-5p inhibits EMT,proliferation and invasion of colon cancer cells by down-regulating EDNRA...