OBJECTIVE: To evaluate the effects and molecular mechanism of action of Changweiqing (CWQ) in reversing multidrug resistance by observing its impacts on nuclear translocation of Y-box binding protein-1 (YB-1), multi-drug resistance gene (MDR1) expression and P-glycoprotein (P-gp) expression in human colon cancer cell line HCT8/V with drug-resistance induced by vincristine. METHODS: Cultured HCT8/V cells were exposed to the CWQ-containing rat serum prepared by drug perfusion. YB-1 expressions in cell plasma and nuclei were examined by Western blot; the binding activity of YB-1 to MDR1 gene promoter sequences was detected by electrophoretic mobility shift assay (EMSA); the mRNA transcription levels of MDR1, YB-1 and multi-resistance related protein (MRP) were examined by RT-PCR; the expression of P-gp on cell membrane was determined by flow cytometry. Results Along with the increasing drug's concentration of CWQ-containing serum from 1.25% up to 2.5% and 5%, the expressions of YB-1 decreased in HCT8/V cell nuclear and increased in cytoplasm gradually; the binding activity of YB-1 to MDR1 gene promoter weakened (P < 0.01), MDR1 mRNA expression and fluorescence intensity of P-gp on cell membrane attenuated (P < 0.05 or P < 0.01), while YB-1 and MRP mRNA unchanged (P > 0.05). CONCLUSION: CWQ could reverse the drug-resistance of colon cancer cells by influencing nuclear translocation of YB-1 and reducing the expression of MDR1/P-gp.
OBJECTIVE: To evaluate the effects and molecular mechanism of action of Changweiqing (CWQ) in reversing multidrug resistance by observing its impacts on nuclear translocation of Y-box binding protein-1 (YB-1), multi-drug resistance gene (MDR1) expression and P-glycoprotein (P-gp) expression in humancolon cancer cell line HCT8/V with drug-resistance induced by vincristine. METHODS: Cultured HCT8/V cells were exposed to the CWQ-containing rat serum prepared by drug perfusion. YB-1 expressions in cell plasma and nuclei were examined by Western blot; the binding activity of YB-1 to MDR1 gene promoter sequences was detected by electrophoretic mobility shift assay (EMSA); the mRNA transcription levels of MDR1, YB-1 and multi-resistance related protein (MRP) were examined by RT-PCR; the expression of P-gp on cell membrane was determined by flow cytometry. Results Along with the increasing drug's concentration of CWQ-containing serum from 1.25% up to 2.5% and 5%, the expressions of YB-1 decreased in HCT8/V cell nuclear and increased in cytoplasm gradually; the binding activity of YB-1 to MDR1 gene promoter weakened (P < 0.01), MDR1 mRNA expression and fluorescence intensity of P-gp on cell membrane attenuated (P < 0.05 or P < 0.01), while YB-1 and MRP mRNA unchanged (P > 0.05). CONCLUSION: CWQ could reverse the drug-resistance of colon cancer cells by influencing nuclear translocation of YB-1 and reducing the expression of MDR1/P-gp.