TRAF6基因沉默对人视网膜母细胞瘤细胞增殖、凋亡和侵袭的影响

Effect of silencing TRAF6 gene on proliferation, apoptosis and invasion of retinoblastoma Y79 cells

目的:探讨体外沉默肿瘤坏死因子受体相关因子6(TRAF6)基因对人视网膜母细胞瘤Y79细胞增殖、凋亡和侵袭的影响。方法:靶向TRAF6基因的小干扰RNA(TRAF6 siRNA)转染Y79细胞,采用qRT-PCR和Western blot法检测转染效果,MTT比色法及细胞克隆实验测定沉默TRAF6对Y79细胞增殖的影响,流式细胞术检测细胞凋亡及细胞周期,Transwell小室检测细胞迁移和侵袭能力。结果:Y79细胞经TRAF6沉默后,TRAF6 mRNA和蛋白表达水平与对照组相比均明显降低,细胞存活率、克隆形成率均明显低于对照组细胞,细胞凋亡率明显高于对照组,同时细胞周期发生明显变化,G0/G1期细胞数目增多,S和G2/M期细胞数目减少,且侵袭细胞数目、迁移细胞数目相比对照组明显减少。结论:沉默TRAF6后能显著抑制视网膜母细胞瘤Y79细胞的生长,促进肿瘤细胞的凋亡,同时抑制其侵袭和迁移能力,TRAF6可能是视网膜母细胞瘤治疗的新靶点。

AIM:To investigate the effect of tumor necrosis factor receptor associated factor 6(TRAF6)on proliferation,apoptosis and invasion of retinoblastoma Y79 cells.METHODS:The Y79 cells were divided into three groups:blank control group,negative control group and TRAF6 siRNA group.After TRAF6 siRNA transfection,the levels of TRAF6 mRNA and protein in Y79 cells were examined by RT-PCR and Western blotting.MTT assay was used to detect cell proliferation.Flow cytometry was employed to detect changes in cell cycle and apoptosis.Cell invasiveness was detected by the Transwell method.RESULTS:Expression of TRAF6 mRNA and protein in the TRAF6 siRNA group significantly decreased compared with the negative and blank control groups.Following the silencing of TRAF6,cell proliferation was inhibited and the apoptosis rate increased;the cell cycle was arrested at G0/G1 phase;the number of cells in S phase was reduced,while the invasion ability of cancer cells decreased.CONCLUSION:Silencing TRAF6 may inhibit the proliferation of Y79 cells,promote cell apoptosis,arrest the cell cycle at G0/G1 phase and decrease the invasive ability.Thus,TRAF6 may be a potential target in therapy for retinoblastoma.