The level of excision repair cross-complementing 1 (ERCC1) gene expression, which is important in the repair of the cisplatin-DNA adducts, is reported to be related to the level of cisplatin resistance in tumor cells. Therefore, ERCC1 is an attractive target to confer increased cellular sensitivity to cisplatin-based chemotherapy. We designed, synthesized, and utilized small interfering RNAs (siRNAs) that were selective for ERCC1 and investigated their effectiveness in altering the repair capacity of the cells to cisplatin-DNA damage as well as the resistance of the cells to cisplatin. Twenty-four and 48h after transfecting ERCC1 siRNA1 and siRNA2 targeting the two different regions of the ERCC1 transcript, both the ERCC1 mRNA and protein expression were significantly inhibited, whereas the mock or control siRNA had no effect. The suppression of ERCC1 expression in the HeLa S3 cells led to a decrease in the repair activity of cisplatin-induced DNA damage along with a decrease in the cell viability against platinum-based drugs, such as cisplatin, carboplatin, and oxaliplatin. A similar increased sensitivity to cisplatin and decreased repair activity were also observed for siRNA-mediated ERCC1 silencing in the MCF-7 and HCT116 cells. This study is the first to demonstrate the feasibility of utilizing ERCC1 siRNAs to specifically reduce the ERCC1 expression level in human cancer cells and provides direct evidence for the potential use of ERCC1 siRNA as a chemotherapy-sensitizing agent.