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目的 探讨外源性p14ARF基因能否对体内外生长的肺癌细胞起抑制增殖的作用。方法 选择 4株具有不同基因背景的肺癌细胞系 ,转染人的野生型p14ARF基因。通过RT PCR、免疫组化及Westernblot杂交检测 ,筛选出同时表达p14ARFmRNA及蛋白的阳性克隆。以母细胞及转染空载体细胞为对照 ,比较它们的增殖、细胞周期分布、裸鼠体内成瘤性以及形态学上的差异。结果 三株具有野生型p5 3基因的肺癌细胞在转染p14ARF基因后 ,细胞周期被显著阻滞于G1或G1/G2期 ,增殖受到抑制。结论 外源性野生型p14ARF基因转染能抑制人肺癌细胞的增殖 ,p14ARF基因是人肺癌基因治疗的理想候选基因。
Objective To investigate whether exogenous p14ARF gene can inhibit proliferation of lung cancer cells grown in vitro and in vivo. Methods Four lung cancer cell lines with different genetic backgrounds were selected and transfected with human wild-type p14ARF gene. Positive clones expressing both p14ARF mRNA and protein were screened by RT PCR, immunohistochemistry and Western blot. The parent cells and transfected empty vector cells were used as controls to compare their proliferation, cell cycle distribution, tumorigenicity in nude mice, and morphological differences. Results After transfection of the p14ARF gene, the three lung cancer cells harboring the wild-type p53 gene were significantly arrested in the G1 or G1/G2 phase and the proliferation was inhibited. Conclusion The transfection of exogenous wild-type p14ARF gene can inhibit the proliferation of human lung cancer cells. p14ARF gene is an ideal candidate gene therapy for human lung cancer.