Qingjie Fuzheng Granules Reverse Multidrug Resistance of Colorectal Cancer Resistant Cells via Inducing Apoptosis and Autophagy
Objective:To investigate the mechanism by which Qingjie Fuzheng Granules(QFG)reverse drug resistance in human colorectal cancer HCT-8/5-FU resistant cells.Methods:HCT-8 cells and HCT-8/5-FU resistant cells were cultured,and the resist-ance index(RI)for different chemotherapeutic drugs and the reversal fold(RF)of QFG were calculated.The cells were divided in-to a blank control group(0 mg/mL)and QFG intervention groups with different concentrations(0.25,0.5,1.0 mg/mL).After 48 hours of intervention,the efflux pump function was assessed by rhodamine-123(Rho123)and doxorubicin(DOX)staining.Cell survival was evaluated by colony formation assay.Apoptosis was detected using Annexin V-FITC/propidium iodide(PI)double stai-ning.Autophagy was analyzed using Cyto-ID staining.Protein expression levels of P-glycoprotein(P-gP),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),selective autophagy receptor protein(p62/SQSTM1),and microtubule-associated protein 1 light chain 3 Ⅱ(LC3-Ⅱ)were measured by Western blot.Results:HCT-8/5-FU cells showed resistance to 5-fluorouracil(5-FU),DOX,and cisplatin(DDP).QFG effectively reversed this drug resistance.Compared with the blank control group,QFG significantly increased the average fluorescence intensity of Rho123,DOX,and Cyto-ID(all P＜0.05),inhibited cell colony formation(P＜0.05),and increased the apoptosis rate(P＜0.05).QFG also reduced the protein expression of P-gP and p62(P＜0.05),de-creased the Bcl-2/Bax ratio(P＜0.05),and promoted the protein expression of LC3-Ⅱ(P＜0.05).Conclusion:QFG can promote apoptosis and autophagy,inhibit the efflux pump function of resistant cells,and thereby reverse multidrug resistance.
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