【摘要】 目的　应用野生型p53及p2 1组成融合基因 ,为口腔鳞癌的基因治疗提供一种新的候选融合基因。方法　用反转录PCR从健康人胚肺细胞 (WI 38)中获得p2 1基因cDNA ,将其与p53融合 ,构建成重组质粒pcDNA p53/p2 1 ,并将其转染至口腔鳞癌细胞Tca81 1 3 ,采用RT PCR和Westernblot证实融合基因的表达 ,通过集落形成实验及3H TdR摄入法检测p53/p2 1融合基因对口腔鳞癌细胞的抑制作用。结果　WI 38细胞中获得一条长约为 50 0bp的条带 ,经测序证实其为p2 1全长编码序列 ,将其与p53构建成融合基因 ,获得融合蛋白p53/p2 1的表达。集落形成实验及3H TdR摄入法观察到p53/p2 1融合基因对口腔鳞癌细胞的生长具有明显的抑制作用 ;经RT PCR和Westernblot证实这是p53/p2 1融合表达的结果。结论　p53/p2 1融合基因可以抑制口腔鳞癌细胞的生长 ,且其抑制细胞生长程度大于p53和p2 1单独作用 ,从而为口腔鳞癌细胞的基因治疗提供了一种新的候选靶基因更多还原

【Abstract】 Objective To study the p53/p21 fusion gene as a potential fusion gene for the gene therapy of human oral squamous cell carcinoma Methods p21 cDNA was obtained from normal human embryonic lung cells by RT PCR, fusing with p53 gene The recombinant plasmid pcDNA p53/p21 was constructed by inserting the p53/p21 fusion gene into eukaryotic expression vector pcDNA3 1 and subsequently transfected into human oral squamous cell carcinoma cell line(Tca8113) with lipofectamine RT PCR and Western blot were used to demonstrate the expression of p53/p21 fusion gene Using clonal formation experiment and 3 H TdR incorporation assay were used to evaluate the clonal formation and proliferation ability of Tca8113 cells Results It was observed that p53/p21 fusion gene could inhibit clonal formation and proliferation of human oral carcinoma RT PCR and Western blot demonstrated that it was the expression of exogenous p53/p21 fusion gene that led to the above results Conclusions Transfection of p53/p21 fusion gene to Tca8113 cells could inhibit the tumor cell proliferation and clone formation in vitro, and make itself a potential fusion gene for the gene therapy of human oral squamous cell carcinoma更多还原