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- Title
miR-15a-5p Regulates Oxaliplatin Resistance in Colorectal Cancer through the Wnt/ β-catenin Pathway.
- Authors
Yanjie HUANG; Tong XU; Luwen WEN; Qi FENG; Jining ZHENG
- Abstract
Objectives: To explore the effects and mechanism of miR-15a-5p on oxaliplatin resistance in colorectal cancer HCT116/L cells. Methods: The expreesion of miR-15a-5p in colorectal cancer sensitive cells HCT116 and resistant cells HCT116/L was detected by RT-qPCR method; the eefect of oxaliplatin on the proliferation of HCT116 and HCT116/L cells was detected by MTT, and its IC and drug resistance fold of HCT116/L cells were calculated; the expreesions of Wnt3a, β-Catenin and P-gp in HCT116 and HCT116/L cells were detected by Western Blot method. HCT-116/L cells were divided into 5 groups: blank control group HCT116/L, control group 1 transfected with miR-15a- 5p mimics NC, experimental group 1 transfected with miR-15a-5p mimics, control group 2 transfected with miR-15a-5p inhibitor NC, and experimental group 2 transfected with miR-15a-5p inhibitor. The expreesion of miR-15a-5p in each group was detected by RT-qPCR method and the transfection efficiency was detected. The eefect of different concentrations of oxaliplatin on the proliferation of cells in each group after trans- fection was detected by MTT and the half inhibitory concentration (50% inhibiting concentration, IC50 ) was calculated. The expressions of Wnt3a, β-catenin and P-gap in each group after transfection were detected by Western Blot method, and the expressions of Wnt3a, β-catenin and MDR1 mRNA in each group after transfection were detected by RT-qPCR method. Results: (i) RT-qPCR results showed that the expressions of miR-15a-5p in HCT116/L cells was (0.16±0.05) significantly lower than that in HCT116 cells ( 8<0.05). ( ii) The IC50 of ox¬aliplatin for HCT-116 cells and HCT116/L cells detected by MTT method were (13.51 ±2.62 ) and (103.08 ±12.29) .g/mL, respectively. The drug resistance index of HCT116/L was 7.63. (iii) Western Blot results showed that the expressions of Wnt3a, β-catenin and P-gp in HCT116/L cells were significantly higher than those in HCT-116 cells ( 8 < 0. 05 ). ( iv) Ater successful transfection, the IC of miR-15a-5p mimics group to oxaliplatin decreased to (40.78 ±2.47) xg/mL by MTT method, and its sensitivity to the drug was significantly improved compared with the control group. Western Blot results showed that the relative expressions of P-gap, Wnt3a and β-catenin were significantly down-regulated (al 8 <0.05); RT-qPCR results showed that the relative expressions of MDR1, Wnt3a and β-catenin mRNA were significant¬ly down-regulated (all 8<0.05). ( v) After successful transfection, the expressions of miR-15a-5p in the miR-15a-5p inhibitor transfection group was (0. 38 ±0. 04) ; MTT results showed that its IC for oxaliplatin was up-regulated to ( 132. 77 ±7. 97) xg/mL, and its sensitivity to chemotherapy drug was significantly lower than that of the control group ; Western Blot results showed that the relative expressions of P-gap, Wnt3a and β-catenin were significantly up-regulated ( aH 8 <0. 05 ) ; RT-qPCR results showed that the relative expressions of MDR1, Wnt3a and β-catenin mRNA were significantly up-regulated ( aH 8 < 0. 05 ). Conclusions : Up-regulation of miR-15 a-5 p expressions can reverse the resistance of HCT116/L cell-line to oxaliplatin. Up-regulation or down-regulation of miR-15a-5p will affect the expressions of P-gap and Wnt β- catenin signaling pathway-related proteins, suggesting that the mechanism of miR-15a-5p reversal of drug resistance may be related to the inhi¬bition of Wnt β-catenin pathway, thereby down-regulating the expressions of P-gap.
- Subjects
COLORECTAL cancer; CATENINS; WNT signal transduction; OXALIPLATIN; WESTERN immunoblotting; DRUG resistance; P-glycoprotein
- Publication
Medicinal Plant, 2022, Vol 13, Issue 3, p9
- ISSN
2152-3924
- Publication type
Article
- DOI
10.19600/j/cnki.issn2152-3924.2022.03.003