ERCC1 siRNA ameliorates drug resistance to cisplatin in gastric carcinoma cell lines

Abstract

The present study examined the effects of cisplatin (DDP) on gastric carcinoma cells by inhibiting the expression of excision repair cross-complementing 1 (ERCC1) using RNA interference (RNAi). mRNA and protein expression of ERCC1 were measured in various gastric carcinoma cell lines using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis. Cells were treated with different concentrations of DDP and the cell viability was measured using an MTT assay. The correlation between the expression of the ERCC1 gene and the resistance to DDP in the cells was determined. The specific ERCC1 small interfering RNA (siRNA) was synthesized and then transfected into SGC-7901/DDP cells. Alterations in intracellular ERCC1 mRNA expression and protein levels were detected using RT-PCR and western blot analysis, the number of apoptotic cells were measured using flow-cytometry and the cell viability was measured using an MTT assay. The gene expression of ERCC1 correlated with the resistance to DDP of the cells. mRNA expression of ERCC1 was significantly reduced 24 h following transfection of ERCC1 siRNA compared with the mock control group. In addition, the number of apoptotic cells was increased and cell viability was significantly decreased in the ERCC1 siRNA-transfected group compared with the mock control group, suggesting that the sensitivity of SGC-7901/DDP cells to DDP had significantly increased. Cells transfected with siRNA1, siRNA2 and siRNA3 were significantly more sensitive to DPP (161, 381 and 249%, respectively) compared with the mock controls (P<0.05). The results of the present study showed that drug resistance to DDP in gastric carcinoma is correlated with increased expression of ERCC1; therefore, inhibition of ERCC1 by siRNA may ameliorate resistance to DDP in gastric carcinoma.