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目的 :研究反义细胞周期蛋白 (cyclinA)基因对舌癌细胞周期的调控作用 ,为cyclinA在基因治疗方面提供理论基础。方法 :采用阳离子脂质体转染方法将已构建成功含人全长cyclinA(1 77kb)的反义真核表达载体pAS_A导入舌癌细胞系 (Tca8113) ,经G418选择性培养液筛选 ,获得阳性克隆细胞 ,通过细胞原位杂交检测cyclinA基因的稳定转染情况。结果 :pAS_A成功转染进入Tca8113,转染后细胞内cyclinAmRNA的转录和蛋白表达明显降低。结论 :反义cyclinA基因稳定有效的在Tca8113细胞系中表达
Objective: To study the role of cyclin A gene in the regulation of tongue cancer cell cycle and to provide a theoretical basis for gene therapy. Methods: Antisense eukaryotic expression vector pAS_A was constructed and transfected into human tongue cancer cell line (Tca8113) with successful construction of full-length cyclin A (1 77kb) by cationic liposome transfection method. The positive cells were screened by G418 selective medium The cells were cloned and the stable transfection of cyclin A gene was detected by in situ hybridization. Results: Transfection of pAS_A into Tca8113 resulted in a significant decrease in the transcription and protein expression of cyclinA mRNA. Conclusion: The antisense cyclin A gene is stably and efficiently expressed in Tca8113 cell line