外源p53对人肺癌细胞生长的抑制

Effects of extraneous wild type p53 on malignant growth of human lung cancer cell line

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 study the effects of extraneous wild type p53 on malignant phenotype of human lung cancer cell line (801 D) with mutant type p53 gene. Methods The human lung cancer line (801 D) with mutant type p53 was selected and characterized with PCR SSCP and DNA sequence analysis. Recombinant plasmid PZiP p53 was constructed, which expressed wild type p53. A transfected cell line, 801 D p53 was obtained after transfering the plasmid into 801 D cell line by gene gun mediated and selected by G418. The wild type p53 and the alteration of malignant phenotype were detected in the transfected cell line by PCR method and cell line culture and heteroplastic transplant study.Results The extraneous wild type p53 gene was detected in 801 D p53 cell line cultured in vitro one year after transfection. The growth of the transfected cell line was inhibited comparing the parent cell line. The colony formation inhibition rate of the cell line was 96% and the tumorigenicity in nude mice was suppressed. The tumor growth significantly slow and the average volume of the transplant tumor was 1/28 of that of the parent cell line. Conclusion Extraneous wild type p53 gene may stably exist in the human lung cancer cell line with mutant type p53 after wild type p53 transfection and suppress the malignant phenotype of the transfected cell line. These results indicate that the recombinant plasmid expressing wild type p53 may be useful for gene therapy of human lung cancer.