microRNA-106b在视网膜母细胞瘤组织中表达的临床意义及其生物学功能研究

目的:研究microRNA-106b(miR-106b)在视网膜母细胞瘤组织中的表达及其临床意义,并探讨其对视网膜母细胞瘤细胞活力、增殖及凋亡的影响.方法:采用qRT-PCR检测33例视网膜母细胞瘤及相应癌旁组织中miR-106b的表达并分析其与不同临床病理特征的相关性.在视网膜母细胞瘤细胞中转染miR-106b类似物或miR-106b抑制物,采用MTT、BrdU-ELISA法及流式细胞仪检测细胞活力、增殖及凋亡,Western blot法检测Rb1蛋白表达变化.结果:miR-106b在视网膜母细胞瘤组织中的表达水平明显高于对应的癌旁组织(P<0.01).miR-106b表达与视神经浸润有关(P<0.05).miR-106b在视网膜母细胞瘤细胞系Y79及HXO-Rb44中的表达水平明显高于其在正常视网膜血管内皮细胞 ACBRI-181中的表达水平(P<0.01).miR-106b抑制物明显降低HXO-Rb44细胞中miR-106b的表达水平(P<0.01),并导致细胞活力降低(P<0.01)、增殖能力下降(P<0.01)及凋亡细胞百分比增加(P<0.01).miR-106b类似物明显增加Y79细胞中miR-106b的表达水平(P<0.01),并导致细胞活力增强(P<0.01)、增殖能力增强(P<0.01)及凋亡细胞百分比增加(P<0.05).同时,降低HXO-Rb44细胞中miR-106b水平显著增高细胞内Rb1蛋白表达水平,增高Y79细胞中miR-106b表达水平,明显降低细胞内Rb1蛋白表达水平(P<0.01).结论:miR-106b在视网膜母细胞瘤组织中表达升高且与恶性临床病理特征相关,miR-106b可增强视网膜母细胞瘤细胞活力、增殖能力,减少细胞凋亡,并可降低细胞内Rb1蛋白表达.miR-106b可能在视网膜细胞瘤的发生、发展中发挥重要作用.

Clinical significance and biological function of microRNA-106b expression in retinoblastoma

Objective:To investigate the clinical significance and biological function of microRNA(miR-106b) expression in retinoblastoma,and effects of miR-106b on the viability,proliferation and apoptosis of retinoblastoma cells.Methods:The expressions of miR-106b in 33 specimens of retinoblastoma tissue and precancerous tissues were detected using qRT-PCR,and the correlation of which with clinicopathologic feature was analyzed. The miR-106b mimics or inhibitors were transfected into the retinoblastoma cells. The viability,proliferation and apoptosis of retinoblastoma cells were detected using MTT,BrdU and flow cytometry,and the expression of Rb1 protein was detected using Western blot.Results:The expression of miR-106b in retinoblastoma tissue was significantly higher than that in precancerous tissue(P<0.01),which was not correlated with optic nerve infiltration(P<0.05).The expressions of miR-106b in retinoblastoma cells Y79 and HXO-Rb44 were significantly higher than that in normal retinal vascular endothelial cells ACBRI-181 (P<0.01).MiR-106b inhibitor could obviously inhibit the expression of miR-106b in HXO-Rb44 cells(P<0.01),and significantly lead to the decreasing of cell viability,proliferation and increasing of percentage of cell apoptosis(P<0.05).The decreasing expression of miR-106b in HXO-Rb44 cells could significantly increase the Rb1 protein expression,and the increasing expression of miR-106b in Y79 cells could significantly decrease the Rb1 protein expression.Conclusions:The expression of miR-106b increases in retinoblastoma tissue,which is correlated with the adverse clinicopathological features of retinoblastoma. MiR-106b can strengthen the viability and proliferation,and reduce the apoptosis of retinoblastoma cells and the expression of Rb1 in retinoblastoma cells.MiR-106b may play a key role in the development and progression of retinoblastoma.