尿刊酸修饰的壳聚糖介导野生型p53基因转染对人肺腺癌细胞系H1299增殖和凋亡的影响
Effects of urocanic acid-modified chitosan-mediated wild-type p53 gene transfection on proliferation and apoptosis of human lung adenocarcinoma cell line H1299
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摘要: 目的 ： 研究尿刊酸修饰的壳聚糖(UAC)介导野生型p53(wt-p53)基因转染对人肺腺癌细胞系H1299(p53基因缺失)的增殖抑制和凋亡诱导作用。 方法 ： UAC介导wt-p53基因转染H1299细胞后，采用荧光显微术、RT-PCR法和Western blot法分别检测细胞内绿色荧光蛋白(GFP)、wt-p53 mRNA和蛋白的表达，并通过MTT法分析基因转染对细胞的生长抑制作用，DAPI染色法和琼脂糖凝胶电泳法评价其对细胞的诱导凋亡作用。 结果 ： UAC介导wt-p53基因转染H1299细胞后，大量细胞表达GFP，细胞内wt-p53 mRNA和蛋白表达水平明显上调，细胞生长受到显著地抑制，凋亡细胞数量明显增加，并有清晰的DNA梯状条带出现。 结论 ： UAC介导wt-p53基因转染能够显著地抑制人肺腺癌细胞系H1299的增殖，并诱导其凋亡。Abstract: Aim ：To investigate the roles of urocanic acid-modified chitosan(UAC)-mediated wild-type p53(wt-p53) gene transfection in inhibiting the proliferation and inducing the apoptosis of human lung adenocarcinoma cell line H1299(p53-null). Methods ：Following UAC-mediated wt-p53 gene transfection in H1299 cells，the expression of green fluorescent protein(GFP)，wt-p53 mRNA and protein was assayed by fluorescence microscopy，RT-PCR and Western blotting，respectively；cell growth inhibition was analyzed by MTT assay；and apoptosis was assessed by DAPI staining and agarose electrophoresis. Results ：UAC-mediated wt-p53 gene transfection in H1299 cells resulted in the appearance of many cells expressing GFP，the notable increases of wt-p53 mRNA and protein expression levels，the significant cellular growth inhibition，the obvious increased proportion of apoptotic cells and the clear DNA ladder. Conclusion ：UAC-mediated wt-p53 gene transfection could significantly inhibit the proliferation and induce the apoptosis in human lung adenocarcinoma cell line H1299.
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