p27^(kip1)基因转染及其对人舌癌细胞增殖的影响

Effect of human p27^(kip1) gene transfection on the proliferation of human tongue squmaous cell carcinoma cells

目的:探讨人p27kip1基因对人舌鳞状细胞癌细胞增殖的影响。方法:从正常组织中克隆人p27kip1基因连接至pcDNA3真核表达载体上,应用脂质体将重组质粒转染舌癌Tca8113细胞,观察外源目的基因在靶细胞中的整合及表达、细胞的生长情况及细胞周期的变化。结果:成功克隆p27kip1cDNA并构建真核表达载体,将其导入Tca8113细胞中,经G418筛选得到稳定表达p27kip1基因的细胞株,其细胞的生长受到抑制,增殖下降。G0/G1期细胞百分数由34.973%增至66.064%,细胞出现了凋亡。结论:克隆并在体外转染Tca8113细胞获得了p27kip1的高表达,抑制舌癌细胞由G1期向S期过渡从而抑制了细胞增殖。

Objective：To investigate the effect of human p27^kip1 gene on the proliferation of human tongue squamous cell carcinoma（TSCC） Tca8113 cells. Methods： Human p27^kip1 gene was cloned from normal tissue and reconstructed into eukaryotic expression vector pcDNA3, the recombinant plasmid was transfected into Tca8113 cells with liposome. The integration and expression of exogenous objective gene in the target cells were examined by PCR and flowcytometry, the growth of transfected cells and non-transfected cells was investigated by MTT assay and flowcytometry. Results：p27^kip1 cDNA was successfully cloned and reconstructed into vector pcDNA3. The gene was trasfected into Tca8113 cells and stable transfection was achieved by screening with G418. After p27^kip1 transfection the cell proliferation was inhibited, proliferation index decreased （ P 〈 0.05 ） , apoptosis cell number increased（ P 〈0.05 ） and Bcl-2 protein expressing cell number decreased（ P 〈 0.05 ）. Conchusion：p27kip1 gene transfection may inhibit the proliferation of TSCC cells.