miR-30a抑制人骨肉瘤细胞143B的迁移、侵袭和活力

目的探讨miR-30a对人骨肉瘤细胞143B侵袭、迁移和细胞活力的影响。方法过表达或抑制miR-30a分别处理人骨肉瘤细胞143B。划痕实验观察细胞划痕愈合能力;Transwell实验检测143B细胞迁移和侵袭能力;MTT实验检测细胞活力;定量PCR实验确认过表达miR-30a腺病毒有效性并且检测RUNX2 mRNA表达;Western blot检测细胞中RUNX2总蛋白表达。结果过表达miR-30a抑制了骨肉瘤细胞143B迁移和侵袭(P0.05),在72 h时,miR-30a明显抑制细胞活力(P0.01);抑制miR-30a的内源性表达后,143B细胞的迁移和侵袭能力增加(P0.05),细胞活力也表现出上升水平(P0.01);同时过表达miR-30a可以抑制RUNX2的蛋白表达,抑制内源性miR-30a后RUNX2蛋白水平表达增加(P0.05)。荧光素酶活性检测,miR-30a可以靶向于RUNX2(P0.01)。结论 miR-30a抑制骨肉瘤细胞143B的迁移、侵袭和活力,其作用可以能是通过抑制RUNX2的表达来实现。

miR-30a suppresses migration,invasion and vitality of human osteosarcoma cell line 143B

Objective To investigate the effect of miR-30a on human osteosarcoma cell 143B migration,invasion and cell viability. Methods 143B cells were infected or transfected with recombinant adenovirus miR-30a (Ad-miR30a) and miR-30a inhibitor, respectively. Wound healing assay was performed to detect the cell healing ability(P<0.05). Cell migration and invasion ability were determined by Transwell assay(P<0.05).The cell viability was analyzed by MTT assay(P<0.01). Real-time quantitative PCR was performed to analyze the expression of RUNX2 mRNA level and confirmed the adenovirus miR-30a which would express in 143B cells.The expression of RUNX2 at protein level was analyzed by Western blot. miR-30a target to RUNX2 was verified by luciferase reported gene assay. Results The ability of migration and invasion was suppressed in osteosarcoma cell 143B by overexpression miR-30a,and the cell viability was also decreased .After the endogenous miR-30a was inhibited, the cell motility and invasion were enhanced, the cell viability was promoted.The RUNX2 protein was decreased after overexpression miR-30a compared with control group in 143B cell. The luciferase activity of RUNX2 was decreased by adding miR-30a.Conclusions 143B cell migration, invasion and viability were suppressed by miR-30a,and this process might be achieved via suppressing RUNX2 protein expression.