miR-142-3p靶向Wnt/β-链蛋白通路对结直肠癌细胞增殖的影响

Effect of miR-142-3p Targeting Wnt/β-catenin Signaling Pathway on Proliferation of Colorectal Cancer Cells

目的探究miR-142-3p靶向Wnt/β-链蛋白(β-catenin)通路对结直肠癌(CRC)细胞增殖的影响。方法选取2018年1月—2020年10月收治的66例CRC的肿瘤组织及其相邻正常组织。同时培养正常人结肠上皮细胞NCM460和CRC细胞系(HT29、LoVo、HCT116、Caco2、SW480细胞)。通过qRT-PCR检测CRC肿瘤组织、正常组织、正常人结肠上皮细胞与CRC细胞系中miR-142-3p表达水平;经免疫蛋白印迹法检测CRC细胞系中β-catenin表达水平;采用CCK-8法和流式细胞术检测miR-142-3p过表达后对CRC细胞增殖的影响;经免疫蛋白印迹检测过表达miR-142-3p对Wnt/β-catenin信号通路相关蛋白的影响;采用双荧光素酶报告基因检验miR-142-3p与β-catenin编码基因CTNNB1的靶向关系。结果miR-142-3p在CRC肿瘤组织和CRC细胞系中的表达显著下降(P<0.05);β-catenin在CRC细胞系中的表达显著升高(P<0.05);过表达miR-142-3p可靶向结合CTNNB1,显著抑制Caco2、LoVo和HT29细胞的增殖和Ki67+细胞比例,抑制β-catenin、c-myc和Cyclin D1的表达(P<0.05)。结论miR-142-3p可通过靶向调节β-catenin的表达,干扰Wnt/β-catenin通路,抑制CRC细胞的增殖。

Objective To explore effect of miR-142-3p targeting Wnt/β-catenin pathway on proliferation of colorectal cancer(CRC)cells.Methods Tumor tissues and adjacent normal tissues of 66 CRC patients admitted during January 2018 and October 2020 were selected.Meanwhile,normal human colon epithelial cells NCM460 and CRC cell lines(HT29,LoVo,HCT116,Caco2 and SW480)were cultured.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect miR-142-3p expressions in CRC tumor tissues,normal tissues,normal human colonic epithelial cell and CRC cell lines.Expressions ofβ-catenin in CRC cell lines was detected by Western blot.Effect of miR-142-3p overexpression on proliferation of CRC cells was detected by CCK-8 method and flow cytometry.Western blot was used to detect effects of miR-142-3p overexpression on Wnt/β-catenin signaling pathway related proteins.Double luciferase reporter genes were used to verify the targeting relationship between miR-142-3p and CTNNB1.Results Expressions of miR-142-3p in CRC tumor tissues and CRC cell line were significantly decreased(P<0.05),while expression ofβ-catenin in CRC cell line was significantly increased(P<0.05).Overexpression of miR-142-3p could target CTNNB1 and significantly inhibit proliferation of Caco2,LoVo and HT29 cells and proportion of Ki67+cells,and inhibit expressions ofβ-catenin,c-myc and Cyclin D1(P<0.05).Conclusion The miR-142-3p may inhibit proliferation of CRC cells by targeting modulation ofβ-catenin expression and interfering Wnt/β-catenin signaling pathway.