miR-21在人骨肉瘤细胞系U2OS体外侵袭中的作用及其机制

Role and mechanism of miR-21 in human osteosarcoma cell line U2OS invasion in vitro

目的:探讨miR-21在人骨肉瘤细胞系U2OS体外侵袭中的作用及其机制。方法:培养人正常成骨细胞系hFOB 1.19和人骨肉瘤细胞系U2OS,通过实时荧光定量PCR(q RT-PCR)比较2组细胞miR-21的表达差异。通过miR-21-up慢病毒感染U2OS细胞,qRT-PCR检测感染前后U2OS细胞中miR-21的表达变化。Transwell侵袭实验检测miR-21对U2OS细胞侵袭的影响,并通过Western blot检测验证在U2OS细胞中miR-21对于靶基因PTEN是否具有调控作用及miR-21对U2OS细胞中侵袭相关蛋白基质金属蛋白酶(MMP)-2、MMP-9表达的影响。结果:q RT-PCR结果表明miR-21在骨肉瘤细胞系U2OS中高表达(P<0.05)。Transwell检测结果表明上调miR-21的表达能够明显促进U2OS细胞的侵袭(P<0.05)。Western blot检测表明PTEN在骨肉瘤细胞中低表达,且与miR-21表达呈负相关(P<0.05)。miR-21可促进侵袭相关蛋白MMP-2、MMP-9的表达(P<0.05)。结论:miR-21可通过抑制PTEN蛋白的表达,进而促进侵袭相关蛋白MMP-2、MMP-9的表达,促进人骨肉瘤细胞系U2OS的侵袭。

Objective： To study the role and mechanism of miR-21 on regulating invasion in osteosarcoma cell line U2OS in vitro. Methods： The cultivation of human fetal osteoblastic cell line hFOB1.19 and human osteosarcoma cell line U2OS. qRT-PCR was used to detect the expression of miR-21 in the previous two groups. miR-21-up lentivirus infected U2OS cells. qRT-PCR was then used to detect the expression of miR-21 between the miR-21-up-infected U2OS cells and no-infected U2OS cells. Transwell chamber invasion assay was used to observe the effect of miR-21 in U2OS cells invasion. Western blot analysis was used to verify miR-21＆#39;s role in regulating the expression of PTEN and invasive associated protein MMP-2/MMP-9 in U2OS cells. Results： qRT-PCR results showed that the expression of miR-21 was significantly increased in U2OS cells （P〈0.05）. Transwell assay results showed that overexpression of miR-21 could significantly promote U2OS cells invasion （P〈0.05）. Western blot analysis results showed that the expression of PTEN was decreased in U2OS cells and negative cor-related with miR-21 （P〈0.05）. Western blot results analysis also showed that miR-21 could significantly improve the expression of MMP-2/9 （P〈0.05）. Conclusion： miR-21, which targets PTEN, promotes U2OS invasion by activating the expression of MMP-2/9.