miR-21在人骨肉瘤细胞系U2OS体外侵袭中的作用及其机制

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摘要目的：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（qRT-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：qRT-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。

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	吕晨
	陈鑫
	朱雄白
	林文军
	王路
	黄正湘
	杨胜武

关键词 ： miR-21, 骨肉瘤, PTEN, 基质金属蛋白酶, 侵袭

Abstract：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’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 correlated 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

Key words： miR-21 osteosarcoma PTEN matrix metalloproteinases invasion

收稿日期: 2016-09-14

基金资助:浙江省自然科学基金资助项目（LY14H060007）

通讯作者: 杨胜武，主任医师，硕士生导师，Email：yangsheng-wu188@sina.com

作者简介: 吕晨（1989-），男，江苏盐城人，住院医师，博士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I7/490

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