Bcl-2短发夹状RNA增强甲氨喋呤对人淋巴瘤裸鼠移植瘤生长的抑制作用  

Small interfering RNA targeting Bcl-2 enhances the inhibitory effects of MTX on transplanted human lymphoma in nude mice

作　　者：房宝英[1] 何冬梅[1] 张洹[1] 刘革修[1] 

机构地区：[1]暨南大学医学院血液病研究所,广州510632

出　　处：《第二军医大学学报》2009年第3期260-264,共5页Academic Journal of Second Military Medical University

基　　金：广东省自然科学基金(04010446)~~

摘　　要：目的:观察Bcl-2短发夹状RNA(short hairpin RNA,shRNA)与甲氨喋呤(methotrexate,MTX)联用后对人淋巴瘤裸鼠移植瘤生长的抑制作用。方法:45只裸鼠注射Raji细胞悬液制备裸鼠人淋巴瘤模型,将聚乙烯亚胺(polyethylenimine,PEI)与Bcl-2shRNA载体质粒(本实验室制备)的混合物及MTX直接注入肿瘤内,观察Bcl-2shRNA及联合MTX对肿瘤生长的影响。在用药21d后无痛处死动物,分离皮下肿瘤,称重,计算抑瘤率;H-E染色观察肿瘤组织的病理形态;以RT-PCR法检测Bcl-2mRNA在肿瘤内的表达情况。结果:Bcl-2shRNA与MTX联用组移植瘤的生长最慢,与单用Bcl-2shRNA和MTX相比差异有统计学意义(P<0.05)。处理结束后剥离瘤块,Bcl-2shRNA组瘤质量与阴性shRNA组、空质粒组相比差异均有统计学意义(P<0.05)。Bcl-2shRNA与MTX联用对裸鼠移植瘤生长的抑制率最高,高于Bcl-2shR-NA和MTX的单用组(P<0.05)。H-E染色可见使用Bcl-2shRNA组及MTX组出现明显的凋亡和组织坏死。瘤组织的单细胞悬液经RT-PCR检测显示Bcl-2mRNA表达在Bcl-2shRNA组均显著下降(P<0.05),而对照组则无明显变化。结论:Bcl-2shRNA可增强MTX对人淋巴瘤裸鼠移植瘤生长的抑制作用。Objective：To explore whether short hairpin RNA （shRNA） targeting Bcl 2 can enhance the inhibitory effect of methotrexate （MTX） on growth of subcutaneously transplanted human lymphoma in nude mice. Methods： Recombinant shRNA expression vector targeting the coding region of Bci-2 mRNA was constructed and preserved in our lab. Human lymphoma Raji ceils were injected subcutaneously into 45 nude mice to establish lymphoma models. The polyethylenimine （PEI）/shRNA complex and （or） MTX were injected into tumors. The influence of Bcl-2 shRNA and （or） MTX on tumor growth was observed. The animals were sacrificed 21 days after administration of drugs and the tumors were removed and weighed; the tumor inhibitory rate was calculated. H E staining was used to observe the pathological morphology of the tumor. The expression of Bei-2 mRNA in the tumor tissues was examined by RT-PCR. Results： The tumor growth was significantly slower in Bcl-2 shRNA/MTX group than in Bcl-2 shRNA or MTX alone groups （P〈0.05）. The tumor weight of mice in Bel-2 shRNA plus MTX group was significantly lower than those in negative shRNA and blank plasmid group （P〈0.05）. The irihibition rate of tumor growth in Bcl-2 shRNA/MTX was significantly higher than those in the Bcl-2 shRNA or MTX alone groups （P〈 0.05）. H E staining showed obvious apoptosis and necrosis in Bcl-2 shRNA group and MTX group. RT PCR result showed that the expression of Bcl-2 mRNA in tumor cell suspension was significantly decreased in Bcl-2 shRNA group （P〈0.05）, and kept unchanged in the control group. Conclusion： The shRNA targeting Bcl-2 can enhance the inhibitory effect of MTX on the growth of subcutaneously transplanted human lymphoma in nude mice.

关 键 词：淋巴瘤 RAJI细胞 BCL-2基因 RNA干扰 甲氨喋呤 

分 类 号：R733.41[医药卫生—肿瘤]