MiR-506 enhances the sensitivity of human colorectal cancer cells to oxaliplatin by suppressing MDR1/P-gp expression.

Abstract

Chemoresistance development represents a major obstacle to the successful treatment of colorectal cancer (CRC). The aim of this study was to elucidate the mechanism by which miR-506 reverses oxaliplatin chemoresistance in CRC. In this study, miR-506 levels were measured in 74 patients with colon cancer via quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH). We subsequently analysed the relationship between miR-506 expression and CRC patient survival via the Kaplan-Meier method. MTT assay demonstrated the fractional survival rates and cell viability of HCT116-OxR, HCT116-OxR-miR-Ctrl and HCT116-OxR-miR-506 cells treated with oxaliplatin at different concentrations. Cell proliferation and apoptosis were assessed via flow cytometry (FCM) analysis and apoptosis assay. MDR1 mRNA expression and P-gp protein expression were assessed via qRT-PCR and Western blotting (WB) respectively. Immunofluorescence (IF) staining demonstrated P-gp expression in HCT116-OxR and HCT116-OxR-miR-506 cells. qRT-PCR and WB were used to detect Wnt/β-catenin pathway activity after miR-506 overexpression. In the present study, in ISH and qRT-PCR results demonstrated that miR-506 is weakly expressed in chemoresistant CRC tissues. The low miR-506 expression group exhibited lower 5-year OS and lower 5-year RFS than the high miR-506 expression group. miR-506 overexpression inhibited cell growth and increased oxaliplatin-induced cell apoptosis in HCT116-OxR cells, as shown via FCM and apoptosis assay. We subsequently noted low MDR1/P-gp expression in HCT116-OxR-miR-506 cells via qRT-PCR, WB and IF. Lastly, we demonstrated low MDR1/P-gp expression in HCT116-OxR-miR-506 cells via inhibition of the Wnt/β-catenin by WB, MTT and FCM analysis. Taken together, the findings of our study demonstrate that miR-506 overexpression in HCT116-OxR cells enhances oxaliplatin sensitivity by inhibiting MDR1/P-gp expression via down-regulation of the Wnt/β-catenin pathway and thus provide a rationale for the development of miRNA-based strategies to reverse oxaliplatin resistance in CRC cells.