YY1基因在肾癌组织中的表达及对肾癌细胞功能的影响  

Expression of YY1 in human renal cell carcinoma and effects of YY1-targeting siRNA on biological features of renal cancer cell line in vitro

作　　者：王明丽[1] 李智[1] 何丽梅[1] 徐晋豫[1] 许闪闪[1] 

机构地区：[1]同济大学附属第十人民医院检验科,上海200072

出　　处：《同济大学学报（医学版）》2012年第4期26-31,共6页Journal of Tongji University(Medical Science)

摘　　要：目的探讨YY1基因在肾癌组织中表达及小干扰RNA(siRNA)对人肾癌786-O细胞增殖和侵袭的影响。方法以Real-Time PCR检测转录因子YY1在20对肾癌及癌旁组织中的表达;采用脂质体转染法将siRNA转染入人肾癌786-O细胞中,Real-Time PCR和Western印迹法检测YY1转染效率;MTT和平板克隆形成实验检测细胞增殖变化;流式细胞术检测细胞周期;Transwell及Matrigel检测细胞侵袭能力,Real-Time PCR检测基质金属蛋白酶MMP9及钙黏附蛋白E-cadherin mRNA表达水平。结果与癌旁组织相比,20对癌组织中YY1 mRNA表达量显著增高(P<0.01)。转染人肾癌786-O后,siRNA-YY1组细胞YY1mRNA的表达抑制率达66%(P<0.01),蛋白表达水平也明显低于siRNA-NC组。与NC组相比,siRNA-YY1组细胞增殖能力显著降低(P<0.01),细胞生长被阻滞在G0/G1期,细胞侵袭能力明显减弱(P<0.01)。此外,与NC组相比,MMP9及E-cadherin mRNA水平有显著差异(P<0.01)。结论 YY1在肾癌组织中高表达,靶向沉默YY1可以在体外显著抑制肿瘤细胞的生长和侵袭能力。YY1特异性siRNA作为一种治疗肿瘤的新途径值得进一步研究。Objective To investigate the expression of YY1 in human renal cell carcinoma and the inhibitory effect of small interfering RNA on the expression of YY1 and the biological features of human renal carcinoma cell line 786-0. Methods The expression of YY1 mRNA levels in cancer and pericancerous tissues were detected by Real-Time PCR. Small interfering RNA was transfected into 786-0 cells, the expression levels of YY1 mRNA and protein were detected by Real-Time PCR and Western blotting. The proliferation inhibition was determined by MTT assay and cloning formation tests. The cell cycle distribution was assessed by flow cytometry. The invasive ability was examined by Transwell and Matrigel invasive assay; the expression levels of MMP9 and E-cadherin mRNA weredetected by Real-Time PCR. Results The YYI mRNA levels in cancer tissues were significantly higher than those in pericancerous tissues（ P 〈 0.01 ）. After transfection, the relative expression level of YY1 mRNA in the YYI-siRNA group was reduced by 66% （ P 〈 0.01 ） as compared to that of NC- siRNA group, and the protein level was also significantly decreased. Compared to NC group, the proliferation of cells in YYI siRNA group was significantly inhibited （ P 〈 0.01 ）, the cell cycle was arrested at G0/G1 phase. Transwell and Matrigel showed that invasive ability of 786-0 cells were significantly suppressed after the transfection（P 〈 0.01 ）. Conclusion YY1 is over expressed in human renal cell carcinoma, YY1 -targeting siRNA can inhibit the proliferation and invasiveness of human renal cancer 786-0 cells.

关 键 词：肾肿瘤 YY1基因 RNA干扰 细胞增殖 侵袭 

分 类 号：R737.11[医药卫生—肿瘤]