凋亡素基因转染对人胆管癌细胞凋亡作用的研究

Apoptosis of human bile duct carcinoma cell induced by Apoptin gene transfection

目的探讨凋亡素对人胆管癌细胞凋亡的影响。方法以PCR方法获得凋亡素基因,然后运用T4连接酶将凋亡素基因和含荧光蛋白基因的质粒pAdtrack-CMV连接,通过酶切和测序证实成功构建重组质粒。再以DOTAP脂质体介导凋亡素基因转染人胆管癌细胞株QBC939。通过TUNEL染色证实凋亡素能否导致人胆管癌细胞的凋亡。结果酶切和测序均证实了成功构建含凋亡素基因和荧光蛋白基因的重组质粒。TUNEL染色显示凋亡素基因的转染引发人胆管癌细胞株QBC939的凋亡率与对照组相比具有非常显著的差异(P<0.01)。结论凋亡素能诱导培养的人胆管癌细胞的凋亡,在胆管癌的基因治疗上有深入研究的价值。

Objective To investigate the apoptosis of human bile duct carcinoma cell induced by Apoptin gene transfection in vitro. Methods Apoptin gene was amplified by polymerase chain reaction （PCR） and then inserted into pAdtrack-CMV containing green fluorescent protein gene through T4 DNA ligase. The reconstructed plasmids were identified by enzyme digestion and sequencing. Apoptin expression vector was transfected into human bile duct carcinoma cell line QBC939. The apoptosis of human bile duct carcinoma cell induced by Apoptin gene was detected by TUNEL. Results Reconstructed pAdtrack-CMV containing Apoptin gene was obtained successfully. TUNEL test showed that there was significant difference in apoptosis index between Apoptin gene transfection and the control group （P 〈 0. 01 ）. Conclusion Apoptin can induce apoptosis of human bile duct carcinoma cell. It may be a promising tool for the gene therapy of cholangiocarcinoma.