Bmi-1基因沉默对人视网膜母细胞瘤SO-RB_(50)细胞生长的体外抑制作用  

Inhibitory Effect of Bmi-1 Knockdown on Growth of Human SO-RB_(50) Cells In Vitro

作　　者：谢满云[1] 钟秀风[1] 李永平[1] 郑健樑[1] 林健贤[1] 张文忻[1] 

机构地区：[1]中山大学中山眼科中心//眼科学国家重点实验室病理室,广东广州510060

出　　处：《中山大学学报（医学科学版）》2011年第6期723-728,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然基金(30672276);广东省科技计划项目(2006B36006007;2008B030301081)

摘　　要：【目的】探讨体外沉默Bmi-1基因对人视网膜母细胞瘤SO-RB50瘤细胞增殖和凋亡的影响。【方法】免疫组织化学和RT-PCR分别检测Bmi-1基因在人视网膜母细胞瘤瘤组织和SO-RB50瘤细胞中的表达。体外化学合成法合成的靶向Bmi-1基因的siRNA双链转染培养的人SO-RB50瘤细胞。荧光定量RT-PCR和western-blot分别检测转染Bmi-1 siRNA后的人SO-RB50瘤细胞中Bmi-1 mRNA和蛋白水平的变化。MTT法测定Bmi-1基因干扰后SO-RB50瘤细胞增殖情况。流式细胞仪检测Bmi-1 siRNA对人SO-RB50瘤细胞凋亡的影响。【结果】免疫组化和RT-PCR显示Bmi-1基因在人视网膜母细胞瘤瘤组织和SO-RB50瘤细胞中表达。人SO-RB50瘤细胞经Bmi-1沉默处理后,与阴性对照组相比,Bmi-1在mRNA和蛋白水平抑制率分别为(83.9±3.2)%和(82.8±1.1)%。MTT结果显示干扰组细胞在第3天增殖抑制作用最明显,抑制率达(52.5±1.9)%。干扰组人SO-RB50瘤细胞凋亡率为(20.7±1.1)%,阴性组细胞凋亡率为(1.9±0.2)%,两者有统计学差异。【结论】Bmi-1特异性siRNA能显著抑制人SO-RB50瘤细胞Bmi-1基因的表达,抑制细胞生长,可能通过促进瘤细胞凋亡而起作用,Bmi-1基因可能为RB治疗的新靶点。[Objective] To investigate the effect of Bmi-1 gene knockdown on the proliferation and apoptosis in human SO-RB5o ceils. [ Methods ] Bmi-1 expression was examined in human retinoblastoma tissue and SO-RBso cells by immunohistoehemistry and RT- PCR. Chemically synthesized Bmi-1 siRNA was transfected into SO-RBso cells. After Bmi-1 siRNA transfection, expression of Bmi-1 mRNA and protein in SO-RB5o cells were examined by real time RT-PCR and Western blot analysis. Cell growth and proliferation were analyzed by MTT assay. Cell apoptosis was observed by flow cytometry. [ Results ] Bmi-1 was expressed in human retinoblastoma tissue and SO-RB50 cells. After Bmi-1 siRNA treatment, Bmi-1 mRNA and protein in SO-RB50 cells were dramatically decreased by （83.9± 3.2） % and （82.8± 1.1 ）%, respectively. After Bmi-1 knockdown, MTr showed that Bmi-1 siRNA could significantly inhibit SO-RBso cell proliferation. The inhibitory effect reached peak at 96 hours [ （52.5 ± 1.9）% ] after transfection. Cell apoptosis rate was promoted up to （20.7 ± 1.1）% in the siRNA treated cells while cell apoptosis rate was （1.9 ±0.2）% in the negative group. [Conclusion] Specific Bmi-1 siRNA could inhibit the expression of Bmi-1 and inhibit human SO-RB50 cell growth in vitro. The effect might be attributed to promoting cell apoptosis. These results suggest that Bmi-1 may be a new therapy target in RBs.

关 键 词：视网膜母细胞瘤:Bmi-1 RNA 干扰 

分 类 号：R739.7[医药卫生—肿瘤]