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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 observe the inhibitory effect of Bcl-2 short hairpin RNA (shRNA) combined with methotrexate (MTX) on the growth of human lymphoma xenografts in nude mice. METHODS: Forty-five nude mice were injected with Raji cell suspension to prepare a model of human lymphoma in nude mice. The mixture of polyethylenimine (PEI) and Bcl-2 shRNA plasmid (prepared in this laboratory) and MTX were directly injected into the tumor. To observe the effects of Bcl-2shRNA and MTX on tumor growth. Animals were euthanized 21 days after treatment, and subcutaneous tumors were isolated and weighed to calculate the tumor inhibition rate. H-E staining was used to observe the pathological changes of the tumor tissues. The expression of Bcl-2 mRNA in the tumors was detected by RT-PCR. Results: The growth of transplanted tumor was the slowest with Bcl-2shRNA combined with MTX. Compared with Bcl-2shRNA and MTX alone, the difference was statistically significant (P <0.05). After the treatment, the mass of tumor mass in Bcl-2 shRNA group was significantly lower than that in negative shRNA group and empty plasmid group (P <0.05). Bcl-2shRNA combined with MTX had the highest inhibitory effect on the growth of xenografts in nude mice, which was higher than that of Bcl-2shR-NA and MTX alone groups (P <0.05). H-E staining showed obvious apoptosis and tissue necrosis in Bcl-2shRNA group and MTX group. The single cell suspension of tumor tissue by RT-PCR showed that the expression of Bcl-2 mRNA in Bcl-2shRNA group was significantly decreased (P <0.05), while the control group had no significant change. Conclusion: Bcl-2 shRNA enhances the inhibitory effect of MTX on the growth of human lymphoma xenografts in nude mice.