miR-490-3p通过抑制TGFβR1基因表达进而抑制结肠癌细胞的转移和侵袭

miR-490-3p inhibits colorectal cancer cell migration and invasion by suppressing TGFβR1 gene expression

目的:研究miR-490-3p在结肠癌细胞(colorectal cance rcell,CRC)转移中的表现和生物功能,及其调控作用机制。方法:通过荧光定量PCR测定miR-490-3p在CRC细胞系的表达水平。细胞转染miR-490-3p以及shmiR-490-3p,观察miR-490-3p的过表达或基因沉默对结肠癌细胞的转移能力是否有影响。miR-490-3p的分子靶标由双荧光素酶报告基因分析和免疫印迹技术进行实验认定。通过划痕实验,Transwell小室基质渗透实验对细胞迁移和侵袭能力进行鉴定。结果:miR-490-3p在CRC细胞系中显著低表达(P<0.05,P<0.01,P<0.001,n>3)。过表达miR-490-3p显著降低CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。miR-490-3p的基因沉默显著增加CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。结肠癌细胞细胞系中过表达miR-490-3p显著降低TGFβR1的基因表达(P<0.001,n>3),miR-490-3p基因沉默显著上调TGFβR1的基因表达(P<0.001,n>3)。过表达miR-490-3p抑制TGFβR1的萤光素酶活性(P<0.001,n>3),miR-490-3p基因沉默促进TGFβR1的萤光素酶活性(P<0.001,n>3)。TGFβR1基因沉默减弱shmiR-490介导的细胞迁移和侵袭促进效应(P<0.01,n>3)。结论:miR-490-3p通过抑制TGFβR1的基因表达从而抑制CRC细胞的转移。

Objective： To investigate the potential role of mi R-490-3p on the regulation of colorectal cancer cell（CRC） migration and invasion, as well as the regulatory mechanism. Methods： The level of mi R-490-3p and TGFβR1 in cells was measured by Real-time PCR. The cell migration and invasion was measured by wound healing experiment and Transwell invasion testing system. The protein level of TGFβR1 was measured by Western blot. The transfection of mi R-490-3p mimics was used to make over-expression of mi R-490-3p in the CRC cells. The transfection of lent-shmi R-490-3p was used to make gene silence of mi R-490-3p in the gastric cells. Results： Gene expression of mi R-490-3p was significantly inhibited in CRC cells（P〈0.05, P〈0.01, P〈0.001, n〉3）. Over-expression of mi R-490-3p significantly inhibited the CRC cell migration and invasion（P〈0.01, n〉3）. Gene silence of mi R-490-3p simulated the CRC cell migration and invasion（P〈0.01, n〉3）. Real-time PCR and western blot results indicated over-expression of mi R-490-3p inhibited TGFβR1 gene expression（P〈0.001, n〉3）, and gene silence of mi R-490-3p stimulated TGFβR1 gene expression（P〈0.001, n〉3）. Over-expression of mi R-490-3p inhibited TGFβR1 derived luciferase activity（P〈0.001, n〉3）, and gene silence of mi R-490-3p stimulated TGFβR1 derived luciferase activity（P〈0.001, n〉3）. The gene silencing of TGFβR1 significantly reduced the shmi R-490 induced promotion of CRC cell migration and invasion（P〈0.01, n〉3）. Conclusion： Mi R-490-3p inhibits colorectal cancer cell migration and invasion by suppressing TGFβR1 gene expression.