| 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-2 shRNA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-2 shRNA及联合MTX对肿瘤生长的影响。在用药21 d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-2 mRNA在肿瘤内的表达情况。结果:Bcl-2 shRNA与MTX联用组移植瘤的生长最慢，与单用Bcl-2 shRNA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-2 shRNA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-2 shRNA与MTX联用对裸鼠移植瘤生长的抑制率最高，高于Bcl-2 shRNA和MTX的单用组(P<0.05)。H-E染色可见使用Bcl-2 shRNA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-2 mRNA表达在Bcl-2 shRNA组均显著下降(P<0.05)，而对照组则无明显变化。结论:Bcl-2 shRNA可增强MTX对人淋巴瘤裸鼠移植瘤生长的抑制作用。 |
| 关键词: 淋巴瘤 Raji细胞 Bcl-2基因 RNA干扰 甲氨喋呤 |
| DOI：10.3724/SP.J.1008.2009.0260 |
| 投稿时间：2008-08-12修订日期：2008-10-10 |
| 基金项目:广东省自然科学基金(04010446). |
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| Small interfering RNA targeting Bcl-2 enhances the inhibitory effects of MTX on transplanted human lymphoma in nude mice |
| FANG Bao-ying, HE Dong-mei*, ZHANG Huan, LIU Ge-xiu |
| (Institute of Hematology, Medical College of Jinan University, Guangzhou 510632, China) |
| Abstract: |
| 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 Bcl-2 mRNA was constructed and preserved in our lab. Human lymphoma Raji cells 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 Bcl-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 Bcl-2 shRNA plus MTX group was significantly lower than those in negative shRNA and blank plasmid group (P<0.05). The inhibition 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. |
| Key words: lymphoma Raji cells Bcl-2 gene RNA interference methotrexate |
