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人p16基因是最近发现的一种肿瘤抑制基因.p16蛋白作为一种细胞增殖的负调控因子,在恶性肿瘤发生发展中起重要作用.为了进一步探讨p16基因的抗肿瘤特性,我们通过运用分子克隆技术构建重组人p16基因表达载体(pcDNA3-p16),并通过电穿孔方法将p16基因导入到人肺腺癌NCI-H460细胞中,经G418筛选获得可稳定表达p16的人肺腺癌细胞,观察p16基因对人肺腺癌细胞周期的影响和细胞增殖的作用.结果表明,导入p16基因能使人肺腺癌细胞生长速度较对照细胞明显减慢,克隆形成率下降,细胞周期G1阻滞.结果证明人p16基因能抑制人肺腺癌细胞的生长.本研究为人肺腺癌p16实验性基因治疗提供了实验依据,说明基于恢复p16肿瘤抑制基因功能的基因治疗在治疗人肺腺癌方面有广阔的应用前景.
Human p16 gene is a newly discovered tumor suppressor gene.p16 protein as a negative regulator of cell proliferation plays an important role in the development of malignant tumor.To further explore the anti-tumor properties of p16 gene, we use molecular cloning Recombinant human p16 gene expression vector (pcDNA3-p16) was constructed and transfected into human lung adenocarcinoma NCI-H460 cell by electroporation. The human lung adenocarcinoma cell line stably expressing p16 was screened by G418. p16 gene on human lung adenocarcinoma cell cycle and cell proliferation.The results showed that the introduction of p16 gene could significantly slow down the growth of human lung adenocarcinoma cells compared with the control cells, the clonogenic rate decreased, G1 cell cycle arrest. The results show that human p16 gene can inhibit the growth of human lung adenocarcinoma cells.This study provides an experimental basis for the experimental gene therapy of human lung adenocarcinoma p16 gene therapy based on the restoration of p16 tumor suppressor gene function in the treatment of human lung adenocarcinoma Broad application prospects.