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目的 生存素 (survivin)基因在胃癌组织中高水平表达 ,而在正常胃黏膜中无表达。生存素表达的水平与胃癌的预后密切相关。通过基因重组构建生存素基因突变体 (pcDNA3 survivin mutant,Cys84Ala)质粒 ,并以脂质体转染的方法将质粒DNA转入胃癌细胞株 ,观察其对胃癌细胞的生物学特性及多药耐药性的影响。方法 应用RT PCR、Westernblot、免疫组化等方法检测胃癌细胞中生存素基因mRNA和蛋白的表达 ,流式细胞仪和吖啶橙染色检测胃癌细胞凋亡。结果 通过RT PCR检测 ,在所有的胃癌细胞株中均有不同程度生存素基因表达 ,生存素基因突变体Cys84Ala通过抑制野生型生存素基因的功能诱导胃癌细胞凋亡 ,增加caspase 3活性和促进细胞色素C的释放 ,增加胃癌细胞对化疗药物的敏感性。结论 突变体Cys84Ala能诱导胃癌细胞凋亡和增加胃癌细胞对化疗药物的敏感性。因此 ,生存素基因突变体Cys84Ala有可能成为胃癌治疗的一个候选基因。
Objective Survivin gene is highly expressed in gastric cancer tissues, but not expressed in normal gastric mucosa. The level of survivin expression is closely related to the prognosis of gastric cancer. To construct the plasmid of survivin gene mutant (Cys84Ala) by gene recombination and transfer the plasmid DNA into the gastric cancer cell line by liposome transfection method to observe its biological characteristics and multidrug resistance in gastric cancer cells Sexual effects. Methods The expressions of survivin mRNA and protein in gastric cancer cells were detected by RT PCR, Western blot and immunohistochemistry. The apoptosis of gastric cancer cells was detected by flow cytometry and acridine orange staining. Results RT-PCR showed that all the gastric cancer cell lines had different levels of survivin gene expression. The survivin gene mutant Cys84Ala induced the apoptosis of gastric cancer cells by inhibiting the function of wild-type survivin gene, increased the activity of caspase 3 and promoted the proliferation of cells Cholesterol C release, increase the sensitivity of gastric cancer cells to chemotherapy drugs. Conclusion Mutant Cys84Ala can induce gastric cancer cell apoptosis and increase the sensitivity of gastric cancer cells to chemotherapeutic drugs. Therefore, survivin gene mutant Cys84Ala may become a candidate gene therapy for gastric cancer.