上调miR-493靶向E2F1抑制骨肉瘤细胞增殖及迁移

摘要

Objective This study investigated the effects of miR-493 on the proliferation and migration of osteosarcoma cells and its possible molecular mechanism.Methods RT-PCR was used to measuremiR-493 expres-sion of osteosarcoma tissue,osteosarcoma cell lines,the control of the tumor tissue and osteoblast cell line. The miR-493 of 143B cells was up-regulated by transient transfection. Cell proliferation was detected by MTT assay. Cell migration was measured by Trans-well method.The miR-493 target gene prediction was done by Targetscan.The protein expression of key gene was detected by Western blotting. Results The relative expression of miR-493 in osteosarcoma tissue and osteosarcoma cell lines were higher than that of the control of the tumor tissue and bone cell.Upregulation of miR-493 inhibited the proliferation and migration of osteosarcoma cell line 143B.The miR-493 targeted the transcription factor E2F1. Upregulation of miR-493 inhibited E2F1and MMP-2,MMP-9 gene protein expression level. Conclusion The miR-493 may inhibit the proliferation and migration of osteosarcoma through target gene E2F1.%目的 探讨miR-493对骨肉瘤细胞增殖及迁移的影响及其分子机制.方法 RT-PCR法分别检测骨肉瘤组织和骨肉瘤细胞系及其对照的瘤旁组织和成骨细胞系miR-493表达水平,瞬时转染法上调143B骨肉瘤细胞的miR-493表达水平,然后采用MTT法检测细胞增殖,Transwell法检测细胞迁移,Tar-getscan在线分析软件预测miR-493靶基因,采用Western blotting检测靶基因及侵袭转移关键基因表达水平变化.结果 miR-493在骨肉瘤组织和骨肉瘤细胞系中的相对表达量分别高于其对照的瘤旁组织和成骨细胞系,上调miR-493抑制了143B细胞的增殖和迁移能力,miR-493靶基因是转录因子E2F1,上调miR-493抑制了骨肉瘤细胞143B的E2F1、MMP-2、MMP-9基因蛋白表达水平.结论 miR-493可能通过靶基因E2F1抑制骨肉瘤增殖和迁移能力.