Study on VP3 Gene Therapy for Murine Transplanted Hepatocarcinoma Delivered by UTMD and PEI-Gal
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摘要: 目的 探讨超声靶向微泡破碎(UTMD)联合半乳糖聚乙烯亚胺(PEI-Gal）介导凋亡素基因对小鼠肝癌移植瘤的抑制作用。方法建立c57BL/6小鼠皮下肝癌移植瘤模型，随机分为4组：对照组；PEI-Gal+质粒组；PEI-Gal+超声+质粒组；PEI-Gal+UTMD+质粒组。对组织行冰冻切片采用免疫荧光法检测转染率、Tunel检测凋亡率。观察肿瘤的体积和组织学变化，同时计算抑瘤率。采用免疫组织化学法检测移植瘤Bcl-2及Caspase-3蛋白在各组肿瘤标本中的表达。结果PEI -Gal+UTMD+pEGFP-N3组基因转染效率明显高于对照组、PEI-Gal+质粒组与PEI-Gal+超声+质粒组(P均<0.01）且对组织无明显损伤。治疗12天后PEI-Gal+UTMD+pCDNA-VP3组肿瘤凋亡率(34.57%±3.56%）、抑瘤率(56.6%)均显著高于对照组和PEI-Gal+质粒组(P均<0.01）。肿瘤组织中Bcl-2蛋白表达明显下降，Caspase-3蛋白表达增加与其他各组比较差异均有统计学意义(P<0.01)。结论 超声靶向微泡破碎联合半乳糖聚乙烯亚胺能显著增强基因转染效率，且对组织无明显影响。联合凋亡素基因能通过诱导凋亡抑制肿瘤生长发挥抗瘤作用。Abstract: Objective To investigate the effects of VTMD and PEI-Gal on VP3 gene mediated apoptosis induction and proliferation suppression in murine transplanted Hepatocarcinoma. Methods Forty mice bearing hepatocarcinoma subcutaneously were divided randomly into 4 groups：control group；PEI-Gal+plasmid group;PEI-Gal+plasmid+ultrasound group and PEI-Gal+UTMD+plasmid group.Histological examination、apoptotic index and transfection efficiency were evaluated by the frozen section.the tumor size was measured regularly.The rate of tumor growth inhibition was calculated and the tumor growth curve was described.The protein expressions of Bcl-2 and Caspase-3 were investigated by immunohistochemistry. Results The highest transfection efficiency was observed in PEI-Gal+UTMD+plasmid group which was significantly higher than that in any other groups(P<0.01)and no tissue damage was seen histologically.The tumor inhibition rate(56.6%）and apoptotic index (34.57%±3.56%）were the highest in PEI-Gal+UTMD+pCDNA-VP3 group compared with those control group and PEI-Gal+pCDNA-VP3 group.The protein expression of Bcl-2 was down-regulated markedly，whereas Caspase-3 was up-regulated remarkedly(P<0.01). Conclusion UTMD associated with PEI-Gal can enhance efficiency of gene transfection significantly.The method associated with VP3 gene can significantly induce apoptosis and inhibit proliferation of transplanted tumors in mice.
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Key words:
- UTMD /
- PEI-Gal /
- VP3 gene /
- murine transplanted Hepatocarcinoma
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