The Construction And Functional Study Of MiR-126Stably Over-expressed Colon Cancer Cell-line

[Background]MicroRNA is a non-coding small RNA molecule, it plays an important role in the carcinogenesis. Based on the previous study of NGX6in colon cancer, we found that miR-126may be associated with metastasis of colon cancer. Studies showed that the miR-126also related with gastric cancer, lung cancer, breast cancer and so on. In our study, the cellular functional experiments were carried out in miR-126stably over-expressed cells. The expressions of target genes from databases prediction were validated. In order to explore the function of miR-126in the development process of colon cancer, provide new targets for colon cancer diagnosis, treatment and prognosis.[Methods](1) MiR-126stably over-expressed SW480cells were established by lentiviral.(2) The cellular functional experiment was carried out in miR-126over-expression cells. The proliferation, migration, invasion of cells were detected through MTT test, the wound&healing test and transwell test, respectively. Cell cycle and cell apoptosis was examined by flow cytometry.(3) The expressions of six target genes(SPRED1、PLK2、CRK、 IRS1、SLC7A5、TOM1) from databases prediction, were validated by real-time PCR in miR-126over-expression SW480cells.[Results](1) The lentiviral infected SW480cells expressed miR-1262.8fold higher than the control group (P<0.05)(2) MTT experiment showed miR-126overexpressed cells’ proliferation significantly decrease during72h and96h. Wound healing test and Transwell result indicated the inhibitory effect of miR-126in celluar migration and invasion. Flow cytometry result showed miR-126would increase the proportion of G1phase and decrease the proportion of S phase, inhibit its proliferation and promote the apoptosis.(3) Real-time PCR validation indicated that the expressions of IRS1、SLC7A5、TOM1decreased in miR-126over-expression group compared to control group(P<0.05), and the expressions of SPRED1、 PLK2、CRK increased(P<0.05).[Conclusion](1) MiR-126lentiviral transfected stably over-expressed SW480cells were established.(2) MiR-126over-expression inhibited the proliferation, migration and invasion of S W480cells and promoted its apoptosis.(3) IRS1、SLC7A5、TOM1were possibly the target genes of miR-126.