miR-30 a抑制人骨肉瘤细胞143 B的迁移、侵袭和活力

摘要

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（P＜0.05），在72 h时， miR-30a明显抑制细胞活力（ P＜0.01）；抑制miR-30a的内源性表达后，143B细胞的迁移和侵袭能力增加（ P＜0.05），细胞活力也表现出上升水平（ P＜0.01）；同时过表达miR-30 a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（P＜0.05）。荧光素酶活性检测，miR-30a可以靶向于RUNX2（P＜0.01）。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。%Objective To investigate the effect of miR-30a on human osteosarcoma cell 143B in migration,invasion andcellviability.Methods 143BcellswereinfectedortransfectedwithrecombinantadenovirusmiR-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 expressed in 143B cells.The expression of RUNX2 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 also decreased .After the endogenous miR-30 a being inhibited , the cell motility and invasion enhanced and the cell viability was promoted .The RUNX2 protein decreased after overexpression miR-30 a as compared with controlgroup.TheluciferaseactivityofRUNX2decreasedbyaddingmiR-30a.Conclusions 143Bcellmigration, invasion and viability were suppressed by miR-30a,and this process is potentially achieved via suppressing RUNX 2 protein expression .