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目的探讨Bcl-2基因特异性的小干涉RNA(siRNA)对食管癌细胞的辐射增敏作用。方法构建表达Bcl-2基因特异性siRNA的真核表达载体,并借助脂质体将其转入食管癌细胞 EC109。Western blot检测Bcl-2蛋白在食管癌细胞的表达,流式细胞仪检测食管癌细胞的凋亡,克隆形成实验和裸鼠移植瘤生长抑制实验检测Bcl-2基因特异性siRNA真核表达载体联合X射线照射对食管癌的生长抑制作用。结果 Bcl-2基因特异性siRNA真核表达载体可抑制Bcl-2蛋白在 EC109细胞中的表达,诱导EC109细胞凋亡,增强X射线照射对EC109细胞克隆形成的抑制能力,增强X射线照射对EC109裸鼠移植瘤生长的抑制作用。结论 Bcl-2基因特异性siRNA真核表达载体可显著增强食管癌细胞对X射线照射的敏感性,具有良好的临床应用前景。
Objective To investigate the radiosensitization effect of Bcl-2 gene-specific small interfering RNA (siRNA) on esophageal cancer cells. Methods The eukaryotic expression vector expressing Bcl-2 gene-specific siRNA was constructed and transfected into EC109 cells by liposome. Western blot was used to detect the expression of Bcl-2 protein in esophageal cancer cells. Flow cytometry was used to detect the apoptosis of esophageal cancer cells. Clonal formation assay and growth inhibition assay in nude mice were used to detect the expression of Bcl-2 gene specific siRNA eukaryotic expression vector X-ray irradiation on the growth inhibition of esophageal cancer. Results Bcl-2 gene-specific siRNA eukaryotic expression vector can inhibit the expression of Bcl-2 protein in EC109 cells, induce apoptosis of EC109 cells, enhance the ability of X-ray irradiation to inhibit the formation of EC109 cells and enhance the effect of X-ray irradiation on EC109 Inhibition of tumor growth in nude mice. Conclusion The specific siRNA expression vector of Bcl-2 gene can significantly enhance the sensitivity of esophageal cancer cells to X-ray irradiation and has a good clinical application prospect.