miR-186通过下调垂体瘤转化基因1的表达抑制骨肉瘤细胞的增殖侵袭并诱导其凋亡

发布时间：2018-01-29 05:54

本文关键词： miR- 骨肉瘤垂体瘤转化基因　出处：《中国肿瘤生物治疗杂志》2017年08期 　论文类型：期刊论文

【摘要】：目的:探讨miR-186对骨肉瘤细胞增殖、凋亡及侵袭能力的影响,并探讨其可能机制。方法:实时荧光定量PCR检测骨肉瘤细胞HOS、U2-OS、Saos-2及成骨细胞NHOst中miR-186表达,并运用人工合成的miR-186模拟片段及对照scramble mimic转染至人骨肉瘤HOS及U2-OS细胞内,运用实时荧光定量PCR检测转染后细胞中miR-186的表达水平;分别运用CCK-8法、流式细胞检测技术及Transwall体外侵袭实验检测过表达miR-186对HOS及U2-OS细胞增殖、凋亡及侵袭能力的影响;运用Western blotting及实时荧光定量PCR检测miR-186过表达对细胞中垂体瘤转化基因1(pituitary tumor transforming gene 1,PTTG1)的蛋白及m RNA表达水平的影响。结果:骨肉瘤细胞中miR-186呈现低表达;转染人工合成的miR-186片段可上调HOS及U2-OS细胞中miR-186的表达;miR-186过表达组细胞的增殖能力较scramble组明显下降(P0.01),转染组HOS[(16.9±2.1)%vs(10.4±1.6)%,P0.05]及U2-OS[(22.6±2.9)%vs(14.1±2.2)%,P0.05]细胞的凋亡比例明显高于scramble组,转染组HOS及U2-OS细胞的穿膜数明显低于scramble组(P0.01);转染组细胞中PTTG1的蛋白及m RNA表达水平均明显下降(P0.01),而scramble组无明显变化(P0.05);结论:miR-186能够抑制骨肉瘤细胞的增殖及侵袭并促进细胞凋亡,其作用机制可能与抑制PTTG1表达相关。
[Abstract]:Objective: to investigate the effect of miR-186 on the proliferation, apoptosis and invasion of osteosarcoma cells and its possible mechanism. MiR-186 was expressed in Saos-2 and NHOst of osteoblasts. Human osteosarcoma HOS and U2-OS cells were transfected with synthetic miR-186 mimic fragment and control scramble mimic. The expression of miR-186 in transfected cells was detected by real-time fluorescence quantitative PCR. CCK-8 assay, flow cytometry and Transwall invasion assay were used to detect the proliferation of HOS and U2-OS cells by overexpression of miR-186. The effect of apoptosis and invasion ability; Western blotting and real-time fluorescence quantitative PCR were used to detect the effect of miR-186 overexpression on pituitary tumor transforming gene 1 (1). Pituitary tumor transforming gene 1. Results: the expression of miR-186 was low in osteosarcoma cells. The expression of miR-186 in HOS and U2-OS cells was up-regulated by transfection of synthetic miR-186 fragments. The proliferative ability of miR-186 overexpression group was significantly lower than that of scramble group, and HOS in transfection group was significantly lower than that in scramble group. [(16.9 卤2.1 vs 10.4 卤1.6) and U2-OS. [The percentage of apoptosis in VSN 14.1 卤2.2mb (P 0.05) was significantly higher than that in scramble group (P < 0.05). The transmembrane number of HOS and U2-OS cells in transfection group was significantly lower than that in scramble group (P 0.01). The expression of PTTG1 protein and m RNA in the transfected cells decreased significantly, while in the scramble group there was no significant change in P0.05. Conclusion: miR-186 can inhibit the proliferation and invasion of osteosarcoma cells and promote the apoptosis of osteosarcoma cells. The mechanism may be related to the inhibition of PTTG1 expression.
【作者单位】：重庆医科大学附属永川医院骨科;南昌大学附属第一医院骨科;
【基金】：国家自然科学基金资助项目(No.81502329) 重庆医科大学附属永川医院院内课题资助项目(No.YCZQN201514)~~
【分类号】：R738.1
【正文快照】： 微小RNA(micro RNAs,mi RNA/mi R)是一类内源HOS、U2-OS及Saos-2细胞株用RPMI 1640完全性、非编码的小RNA分子。通过与靶基因的3’-UTR培养基(含有10%胎牛血清、100 U/ml青霉素和100区完全或不完全的结合,mi RNAs能够参与靶基因的μg/ml链霉素),5%CO2及90%饱和湿度的条件下培

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