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目的研究反义tankyrase1逆转录病毒对舌癌细胞系TCCA-8113的端粒酶活性调节及对细胞生长的抑制作用,探讨以tankyrase1为靶点的舌癌基因治疗的可能性。方法构建含反义tankyrase1的逆转录病毒并转染包装细胞PT67后获得反义重组病毒,感染舌癌TCCA-8113细胞。采用RT-PCR检测tankyrase1表达,端粒酶重复扩增法(telomerase repeat amplification protocol,TRAP)检测端粒酶活性,绘制生长曲线了解细胞生长状态,倒置显微镜观察和流式细胞仪检测细胞凋亡。结果反义tankyrase1逆转录病毒作用后的舌癌细胞tankyrase1表达下降,端粒酶活性及细胞生长受到明显抑制,细胞出现凋亡。结论反义tankyrase1对舌癌TCCA-8113细胞的生长和端粒酶活性具有明显的抑制作用,有可能以tankyrase1为靶点对舌癌进行基因治疗。
Objective To investigate the regulation of telomerase activity and cell growth inhibition by antisense tankyrase1 retrovirus in tongue cancer cell line TCCA-8113 and to explore the possibility of gene therapy of tongue cancer with tankyrase1 as target. Methods The retrovirus containing antisense tankyrase1 was constructed and transfected into the packaging cell PT67 to obtain the antisense recombinant virus and to infect tongue cancer TCCA-8113 cells. The expression of tankyrase1 was detected by RT-PCR, the telomerase repeat amplification protocol (TRAP) was used to detect the telomerase activity, and the growth curve was drawn to observe the cell growth status. The apoptosis was detected by inverted microscope and flow cytometry. Results The antisense tankyrase1 retrovirus inhibited the expression of tankyrase1 in tongue cancer cells. The telomerase activity and cell growth were significantly inhibited, and the cells showed apoptosis. CONCLUSION: Antisense tankyrase1 can significantly inhibit the growth and telomerase activity of tongue cancer TCCA-8113 cells. It is possible to use gene therapy of tankyrase1 to target tongue cancer.