ERCC1和BRCA2基因共沉默有效逆转耐药肺癌细胞对顺铂的耐药性

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Abstract Objective: To investigate the effect of cosilencing of ERCC1 and BRCA2 genes on reversing cisplatin resistance in resistant lung cancer cell A549/DDP. Methods: Small interference RNAs (siRNAs) targeting ERCC1 and BRCA2 genes (ERCC1 and BRCA2siRNA) were transfected into A549/DDP cells using lipofectamine 2000. Transfection efficacy was verified by protein expressions of ERCC1 and BRCA2 detected with Western blotting. BRCA1 and RAD51 proteins expressions as well as the FANCD2 monoubiquitination induced by cisplatin were determined by Western blotting. The cell proliferation and apoptosis rate in A549/DDP cells pre and posttreatment with cisplatin were measured by CCK8 assay and flow cytometry using Annexin V/PI strain, respectively, before and after transfection. The formation of FANCD2 and RAD51 nuclear foci were detected using immunofluorescence staining. Results: Cisplatininduced FANCD2 monoubiquitination and FANCD2 nuclear loci formation were significantly decreased after transfection of ERCC1siRNA. The expressions of BRCA1 and RAD51 proteins and RAD51 nuclear foci induced by cisplatin were markedly reduced after transfection of BRCA2siRNA (P<0.05). Individual transfection of ERCC1siRNA or BRCA2siRNA suppressed cell proliferation and promoted cell apoptosis produced by cisplatin. Cotransfection of the two siRNAs further enhanced the cytotoxicity of cisplatin to A549/DDP cells, as compared to transfection of ERCC1siRNA or BRCA2siRNA alone. Conclusion: Silencing of ERCC1 and BRCA2 gene could reverse the resistance of resistant lung cancer cells to cisplatin through inhibition of the FA pathway and the HR pathway respectively, and the reversion effect of cisplatin resistance was further potentiated by cosilencing of ERCC1 and BRCA2 genes.

Received: 20 November 2016

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http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2017/V27/I01/40

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