Bcl-2基因特异性小干涉RNA增强食管癌细胞辐射敏感性的实验研究
Small interfering RNA specific to Bcl-2 enhances radiosensitivity of oesophageal cancer cells
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摘要: 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。Abstract: Objective To explore the effects of smalf interfering RNA(siRNA) specific to Bcl-2 gene on radiosensitivity of oesophageal cancer cells.Methods The eukaryotic expression vector of Bcl-2 gene siRNA was constructed and introduced into oesophageal cancer cells EC 109 by lipofectamine. Bcl-2 protein expression in EC 109 cells was detected by Western blot. Flow cytometry was employed to evaluate apoptosis of EC109. Clone forming assay and nude mice xenograft assay were used to determine the inhibitory effects of X radiation combined with Bcl-2 gene specific siRNA.Results Western blot analysis demonstrated that eukaryotic expression vector of Bcl-2 gene siRNA successfully suppressed Bcl-2 protein expression in EC 109 cells. Bcl-2 gene siRNA could induce apoptosis of EC 109 cells, enhance the inhibitory effects of X radiation on clone forming ability of EC 109 cells and xenograft growth in nude mice.Conclusion The eukaryotic expression vector of Bcl-2 gene siRNA could enhance the radiosensitivity of oesophageal cancer cells and can be used as a powerful adjunct to conventional radiotherapy.
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Key words:
- Esophageal neoplasma /
- Gene,Bcl-2 /
- Radiosensitivity /
- RNA interference
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