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为研究野生型p53基因对肺腺癌细胞株GLC-82细胞生长的作用,确立腺病毒介导的野生型p53基因在肿瘤治疗中的意义,应用分子克隆技术首先构建了野生型p53复制缺陷型腺病毒重组体,应用生化染色方法确定了重组体的转染效率,以免疫组化法及PCR技术分别确定了腺病毒载体介导的p53基因转染GLC-82细胞后p53蛋白和p53 cDNA的表达情况;最后应用细胞生物学方法检测了p53对GLC-82细胞扩增及细胞周期的影响.结果表明所构建的重组体可以高效、快速转染GLC82细胞,抑制GLC-82细胞扩增,使细胞合成期和分裂后期的量减少,出现细胞凋亡现象:提示野生型p53基因导入可作为治疗肺腺癌的途径之一
To study the effect of wild-type p53 gene on the growth of lung adenocarcinoma cell line GLC-82, we established the significance of adenovirus-mediated wild-type p53 gene therapy in cancer treatment. We first constructed a wild-type p53 replication-defective model by molecular cloning. Recombinant adenoviruses were used to determine the transfection efficiency of the recombinants by biochemical staining. The adenoviral vector-mediated p53 and p53 cDNA transfections in GLC-82 cells were determined by immunohistochemistry and PCR techniques, respectively. Expression conditions; The final application of cell biology methods to detect the effect of p53 on GLC-82 cell proliferation and cell cycle. The results show that the recombinant can be constructed to efficiently and rapidly transfect GLC82 cells and inhibit the expansion of GLC-82 cells. The amount of cell synthesis and anaphase is reduced and apoptosis occurs: It is suggested that the introduction of wild-type p53 gene can be used as a treatment for lung adenocarcinoma