β-葡萄糖醛酸酶前药对βG基因转染人胆管癌细胞的作用

The effect of β-G prodrug on β-glucuronidiase-cDNA transfected bile duct carcinoma cell line QBC939

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 。

Objective To investigate the growth inhibitory effect of β G gene and β G prodrug on human bile duct carcinoma cell line (QBC939). Methods Retroviral vector was used to transfect the pDOR βG gene into the bile duct carcinoma cell line (QBC939) by cation lipofectin. The expression of pDOR βG gene was detected by immunohistochemistry and the transfected cells were also observed. The expression of pDOR βG gene and effect of β G prodrug on transfected cells growth were studied. Results The positive cell clone selected by G418 system was proved to be a bile duct carcinoma model with high expression of βG. The expression of pDOR βG in the transfected cells was first verified by immunohistochemistry, immunoflurescence and in situ hybridization. It was found that β G prodrug had a significantly inhibitory effect on pDOR βG QBC939 cell lines by using MTT method. When the β G prodrug concentration was 0.292 g/L, inhibitory rate of pDOR βG QBC939 cell lines was 67.6%. Conclusion β G gene/β G prodrug system was a new suicide gene system, which may provide a novel gene therapeutic strategy for bile duct carcinoma.