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目的:用RNA干扰技术下调肺癌细胞系A549切除修复交叉互补基因(ER-CC1)的表达,以提高肺癌细胞对化学药物的敏感性。方法:首先构建针对ERCC1基因的SiRNA真核表达载体;其次将载体转染肺癌细胞A549,采用RT-PCR和蛋白质印迹法技术证实转染细胞中ERCC1基因沉默效果良好;最后采用MTT法检测构建真核表达载体转染细胞对顺铂药物毒副反应的影响。结果:成功构建针对ERCC1基因的siRNA真核表达载体;将其转染肺癌细胞A549,可以抑制细胞中ERCC1基因表达,转染细胞较亲本细胞ERCC1基因表达降低48.3%,P<0.05;RNA干扰的A549细胞较亲本细胞对顺铂的敏感性升高了2.93倍。结论:RNA干扰是一种沉默ER-CC1表达的理想方法。沉默ERCC1表达可以提高肺癌细胞的化疗敏感性,它可能是一种有潜质的肺癌辅助治疗新方法。
OBJECTIVE: To down-regulate the expression of ER-CC1 in lung cancer cell line A549 by RNA interference to improve the chemosensitivity of lung cancer cells. Methods: The eukaryotic expression vector of SiRNA targeting ERCC1 gene was constructed. Secondly, the vector was transfected into A549 lung cancer cells and the silencing of ERCC1 gene was confirmed by RT-PCR and Western blotting. Finally, Effect of Nuclear Expression Vector Transfected Cells on Cisplatin Drug Toxicity. Results: The eukaryotic expression vector of siRNA targeting ERCC1 gene was successfully constructed. The transfected A549 cells transfected with A549 cells could inhibit the expression of ERCC1 gene and the expression of ERCC1 gene in transfected cells by 48.3%, P <0.05. A549 cells than the parent cell sensitivity to cisplatin 2.93 times. Conclusion: RNA interference is an ideal method to silence the expression of ER-CC1. Silencing ERCC1 expression can enhance chemosensitivity of lung cancer cells, which may be a potential new adjunct to lung cancer therapy.