miR-375下调YAP1对结直肠癌细胞增殖和迁移的影响

Effects of miR-375 downregulation by inhibiting YAP1 on proliferation and migration of colorectal cancer cells

目的探讨miR-375靶向调控Yes相关蛋白(YAP)的表达对结直肠癌细胞增殖和迁移能力的影响。方法选取2018年1月至2019年6月间兰州市第一人民医院收治的行手术切除的37例结直肠癌患者的癌组织及对应的癌旁组织标本、人结直肠癌细胞系HCT116、NCI-H716、HRC-99和正常结直肠上皮细胞系FHC,采用RT-qPCR分析比较结直肠癌组织与对应癌旁组织及结直肠癌细胞系与FHC细胞中miR-375的表达水平。在细胞系水平,应用CCK-8实验、Transwell迁移实验检测过表达miR-375对结直肠癌细胞增殖及迁移能力的影响,应用Western blot检测过表达miR-375的HCT116细胞中YAP1的表达变化。结果在结直肠癌组织中,miR-375的相对表达量低于癌旁组织,YAP1的相对表达量高于癌旁组织。在所有检测结直肠癌细胞系中,HCT116细胞中miR-375的表达水平最低,过表达miR-375后,HCT116细胞的增殖、迁移能力减弱,YAP1表达水平降低。数据库分析显示,YAP1可能是miR-375下游调控的靶基因,且二者在结直肠癌中的表达呈负相关。结论miR-375可能通过抑制YAP1的表达在结直肠癌中发挥抑癌作用。

Objective Objective To investigate the effects of miR-375 on the proliferation and migration of colorectal cancer cells through regulating the expression of Yes-associated protein(YAP).Methods Colorectal cancer and paracancer tissue specimens,HCT116,NCI-H716 and HRC-99 human colon cancer cell line,and FHC normal colorectal epithelial cells line were collected from 37 patients with colorectal cancer undergoing surgical resection at Lanzhou First People’s Hospital from January 2018 to June 2019.RT-qPCR was used to detect the expression level of miR-375 in colorectal cancer and paracancer tissues as well as in human colon cancer cell line and human colon cancer cell line,and FHC normal colorectal epithelial cells line.For cells lines,CCK-8 assay and Transwell assay were used to detect the effect of miR-375 on proliferation and migration of CRC cells.Transwell assay was used to detect the effect of miR-375 on the migration ability of colorectal cancer cells.Western blot was used to detect the expression of Yap1 in miR-375-overexpressing HCT116 cells.Results That the relative expression of miR-375 in colorectal cancer was lower than in paracancer tissues,while the relative expression of Yap1 was higher than in paracancer tissues.The expression level of miR-375 was the lowest in HCT116 cells among all colorectal cancer cell lines.After overexpression of miR-375,the proliferation and migration ability of HCT116 cells decreased,and the expression level of Yap1 decreased significantly.Database analysis showed that YAP1 might be the downstream target gene of miR-375,and there was a negative relationship between the expressions of these two indices in colorectal cancer.Conclusion miR-375 can suppress colorectal tumor by inhibiting YAP1.