Li Weiling, Li Chaoying, Xiong Qi, Tian Xiang, Ru Qin
Wuhan Institutes of Biomedical Sciences, Jianghan University, Wuhan, Hubei 430056, P.R. China.
Exp Ther Med. 2019 Jun;17(6):4577-4585. doi: 10.3892/etm.2019.7506. Epub 2019 Apr 19.
MicroRNA-10b (miR-10b) has been reported to be specifically upregulated in glioma tissues and cell lines. The aim of the present study was to investigate the effect of miR-10b-5p on the proliferation and invasion of glioma cells, and the possible underlying molecular mechanism. Cell viability was evaluated using an MTT assay, and flow cytometry was performed for cell cycle analysis. The effects of miR-10b-5p on glioma cell migration and invasion were assessed using wound healing and Transwell assays, respectively. The activity of matrix metalloproteinase 2 (MMP2) was also determined using zymography. Furthermore, homeobox B3 (HOXB3) mRNA expression in glioma cells was examined using quantitative polymerase chain reaction analysis. The protein expression levels of HOXB3, high mobility group box 1 (HMGB1) and Ras homolog family member C (RhoC) were further measured using western blotting. It was observed that glioma cells transfected with miR-10b-5p inhibitor exhibited significantly decreased proliferation. The wound healing and Transwell assays demonstrated that the miR-10b-5p inhibitor reduced the ability of glioma cells to migrate and invade, while transfection with miR-10b-5p mimic exhibited the opposite effect. HOXB3 was downregulated by miR-10b-5p at both the mRNA and protein levels. In addition, the expression of proteins associated with migration and invasion, including HMGB1, RhoC and MMP2, was upregulated in glioma cells transfected with miR-10b-5p mimic, while these proteins were downregulated in cells transfected with miR-10b-5p inhibitor. Taken together, the findings of the present study indicated that miR-10b-5p downregulation suppressed glioma cell proliferation and invasion, possibly by modulating HOXB3, which may provide a novel bio-target for glioma therapy.
据报道,微小RNA-10b(miR-10b)在胶质瘤组织和细胞系中特异性上调。本研究旨在探讨miR-10b-5p对胶质瘤细胞增殖和侵袭的影响及其可能的潜在分子机制。采用MTT法评估细胞活力,并进行流式细胞术分析细胞周期。分别采用伤口愈合试验和Transwell试验评估miR-10b-5p对胶质瘤细胞迁移和侵袭的影响。还采用酶谱法测定基质金属蛋白酶2(MMP2)的活性。此外,采用定量聚合酶链反应分析检测胶质瘤细胞中同源盒B3(HOXB3)mRNA的表达。进一步采用蛋白质印迹法检测HOXB3、高迁移率族蛋白B1(HMGB1)和Ras同源家族成员C(RhoC)的蛋白表达水平。结果观察到,转染miR-10b-5p抑制剂的胶质瘤细胞增殖明显降低。伤口愈合试验和Transwell试验表明,miR-10b-5p抑制剂降低了胶质瘤细胞的迁移和侵袭能力,而转染miR-10b-5p模拟物则表现出相反的效果。miR-10b-5p在mRNA和蛋白水平均下调HOXB3。此外,在转染miR-10b-5p模拟物的胶质瘤细胞中,与迁移和侵袭相关的蛋白,包括HMGB1、RhoC和MMP2的表达上调,而在转染miR-10b-5p抑制剂的细胞中这些蛋白表达下调。综上所述,本研究结果表明,miR-10b-5p下调可能通过调节HOXB3抑制胶质瘤细胞的增殖和侵袭,这可能为胶质瘤治疗提供一种新的生物靶点。