微小RNA-3175对胶质瘤生长与侵袭的作用

Effect of microRNA-3175 on growth and invasion of glioma

目的 探讨微小RNA（miR）-3175在胶质瘤中的表达及对胶质瘤生长与侵袭的作用.方法 通过定量反转录聚合酶链反应（qRT-PCR）检测40例人胶质瘤组织和胶质瘤细胞A172、U87中miR-3175的表达水平;用Lipofectamine 2000对胶质瘤细胞转染miR-3175抑制物后,qRT-PCR检测miR-3175的抑制程度,并通过噻唑蓝实验、流式细胞术、Transwell实验分别检测miR-3175对胶质瘤细胞增殖、周期、凋亡、侵袭的作用.Western blot分析miR-3175与凋亡相关蛋白p53、Bcl-2、细胞色素C、Caspase-3的关系.结果 miR-3175在胶质瘤中表达水平明显升高（与对照组比较P＜0.05）.转染miR-3175抑制物后,胶质瘤细胞miR-3175的表达量明显下降（P＜0.05）.miR-3175抑制物可有效抑制胶质瘤细胞增殖和侵袭能力,A172、U87增殖能力分别下降（23.56±8.06）％、（40.33±12.94）％;侵袭细胞数减少（47.83±4.33）％、（45.13±5.03）％;同时miR-3175抑制物可明显提升胶质瘤细胞G0/G1期的比例和凋亡率,A172、U87细胞G0/G1期比例分别升高13.51％、13.22％;细胞凋亡率分别提高（8.85±2.51）％、（12.31±5.20）％.miR-3175表达下降可导致凋亡相关蛋白p53、Bcl-2、细胞色素C、Caspase-3的异常表达（P＜0.05）.结论 miR-3175在人胶质瘤中高表达,miR-3175表达量下降可有效抑制胶质瘤细胞生长和侵袭,促进凋亡.

Objective To investigate the expression of microRNA （miR） -3175 in glioma and its effect on the growth and invasion of glioma. Methods The expression levels of miR-3175 in 40 human glioma tissues and glioma cell A172 and U87 were detected by quantitative reverse transcription polymerase chain reaction （qRT-PCR）. After glioma cells transfection of miR-3175 inhibitor with Lipofectamine 2000, qRT-PCR detected the degree of inhibition of miR-3175. The effects of miR-3175 on glioma cell proliferation, cell cycle, apoptosis, and invasion were detected by MTT assay, flow cytometry assay and Transwell assay, respectively. Western blot was used to analyze the relationship between miR-3175 and apoptosis-related protein p53, Bcl-2, cytochrome C, and Caspase-3. Results The expression level of miR-3175 in glioma was increased obviously （compared with the control group P 〈 0.05 ）. The expression level of miR-3175 was decreased significantly after transfection of miR-3175 inhibitom （P 〈 0.05）. The miR-3175 inhibitors could effectively inhibit the proliferation and invasion abilities of glioma cells. The proliferation ability of U87 and A172 decreased 23.56 ± 8.06% and 40.33 ± 12.94% respectively; the number of the invasive cells reduced 47.83 ±4.33% and 45.13±5.03% respectively; at the same time, the MiR-3175 inhibitors could obviously increase the ratio and apoptosis rate of glioma cell G0/G1 phase, the ratio of A172 and U87 cell G0/G1 phase increased 13.51% and 13.22% respectively; the apoptosis rate increased 8.85 ±2.51% and 12.31 ±5.20% respectively. The decreased miR-3175 expression resulted in the abnormal expression of apoptosis related protein p53, Bcl-2, eytochrome C, and Caspase-3 （P 〈 0.05）. Conclusion The miR-3175 expressed highly in human glioma. The decreased expression level may effectively inhibit the growth and invasion of glioma cells, and promote apoptosis.