Hui Zhou1,2, Changwei Lin1,3, Yi Zhang1, Xiuzhong Zhang1, Chong Zhang1, Pengbo Zhang1, Xingwang Xie2, Zeqiang Ren1. 1. Department of Oncological Surgery, Affiliated Hospital of Xuzhou Medical University, Xuzhou, China. 2. Department of General Surgery, Wuhan Third Hospital, Wuhan, China. 3. Department of Gastrointestinal Surgery, Third Xiangya Hospital, Central South University, Changsha, China.
Abstract
OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
Authors: Jin Lu Tong; Chen Peng Zhang; Fang Nie; Xi Tao Xu; Ming Ming Zhu; Shu Dong Xiao; Zhi Hua Ran Journal: FEBS Lett Date: 2011-10-22 Impact factor: 4.124
Authors: Bei Xie; Linjing Li; Zhewen Zhang; Lei Zhao; Juan Cheng; Cunmin Zhou; Jie Cheng; Jing Yan; Jing Chen; Juan Yi; Bei Wang; Suya Jin; Hulai Wei Journal: J Cancer Date: 2021-05-13 Impact factor: 4.207