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目的 观察外源性野生型p53对有p53基因突变的人肺癌细胞系生长的影响。方法 用PCR SSCP及DNA测序 ,选择p53突变的人肺巨细胞癌系 80 1 D。构建野生型p53表达质粒PZiP p53。用基因枪介导外源基因。经G41 8筛选得到转染细胞系 80 1 D p53。用PCR检测外源基因 ,观察转染细胞恶性生长的变化。结果 转染细胞系 80 1 D p53体外长期传代有外源性p53基因存在 ,转染细胞生长明显受到抑制 ,集落形成抑制率达 96% ,裸鼠异种移植致瘤性降低 ,肿瘤生长明显缓慢。结论 外源性野生型p53经基因枪导入有p53基因突变的人肺癌细胞后可长期存在于转染细胞中 ,且明显抑制所转染的癌细胞的恶性生长。
Objective To observe the effect of exogenous wild-type p53 on the growth of human lung cancer cell lines with p53 gene mutation. Methods The p53 mutant human lung giant cell carcinoma line 80 1 D was selected by PCR and DNA sequencing. The wild type p53 expression plasmid PZiP p53 was constructed. Gene guns are used to mediate foreign genes. The transfected cell line 80 1 D p53 was screened by G41 8 . Exogenous genes were detected by PCR to observe the malignant growth of transfected cells. Results The transfected cell line 80 1 D p53 had exogenous p53 gene in vitro for a long time. The growth of transfected cells was obviously inhibited. The inhibition rate of colony formation was 96%. The tumorigenicity of xenografts in nude mice was decreased and the tumor growth was slow. Conclusion Exogenous wild-type p53 can be permanently transfected into human lung cancer cells with p53 gene mutation introduced by gene gun, and it can significantly inhibit the malignant growth of the transfected cancer cells.