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目的 探讨 βG基因 /β 葡萄糖醛酸苷酶 (β G)前体药系统对人胆管癌细胞株QBC93 9的抑制作用。方法 构建真核表达的逆转录病毒载体pDOR βG ,利用阳离子脂质体将其转入人胆管癌细胞株QBC93 9,检测其表达及 β G前体药对转基因细胞的增殖阻断作用。 结果 经G418筛选后的阳性细胞克隆 ,为高表达 βG的胆管癌模型。免疫组织化学、免疫荧光、原位杂交等证实了pDOR βG在转基因细胞中的表达 ;四唑蓝 (MTT)比色试验测定 β G前体药对QBC93 9 pDOR βG细胞有显著的抑制增殖作用 ,β G前体药浓度为 0 .2 92 g/L时 ,其抑制率为 67.6%。QBC93 9 βG组与对照组比较差异有非常显著性 (P <0 .0 1)。结论 βG基因 /β G前体药系统是一种新的自杀基因系统 ,可作为胆道肿瘤基因治疗的一种新方法
Objective To investigate the inhibitory effect of βG gene/β-glucuronidase (β G) prodrug system on human cholangiocarcinoma cell line QBC93 9. Methods The eukaryotic expression retroviral vector pDOR βG was constructed and transfected into human cholangiocarcinoma cell line QBC93 using cationic liposomes. The expression of the recombinant plasmid pDOR βG was detected and the proliferative blocking effect of β G prodrug on transgenic cells was also investigated. Results The positive cell clones screened by G418 were high-expression βG cholangiocarcinoma models. Immunohistochemistry, immunofluorescence, and in situ hybridization confirmed the expression of pDOR βG in transgenic cells, and tetrazolium (MTT) colorimetric assay showed that β G prodrugs significantly inhibited proliferation of QBC93 9 pDOR βG cells. When the concentration of β G prodrug was 0.22 g/L, the inhibition rate was 67.6%. The difference between QBC93 9βG group and control group was very significant (P < 0.01). Conclusion βG gene /β G prodrug system is a new suicide gene system and can be used as a novel method for gene therapy of biliary tumors.