Curcumin Reversal The Resistance Of Oxaliplatin Resistant Human Colon Carcinoma Cell Line HCT-116/L-OHP

Objective:To construct oxaliplatin(L-OHP)drug-resistant cell line HCT-116/L-OHP in human colon cancer.To observe the reversal effect of curcumin(cur)on its drug resistance,and preliminarily explore the possible drug resistance mechanism.Methods:The concentration gradient increasing method was used to gradually increase the L-OHP concentration of HCT-116 in parental colon cancer cells,and the cell line HCT-116/L-OHP resistant to L-OHP was established.The cytotoxicity of L-OHP and curcumin to HCT-116 and HCT-116/L-OHP cells was detected by cck-8 method to observe whether curative resistance could be reversed.Flow cytometry was used to detect apoptosis rate and cell cycle Western blot was used to detect the expression of drug-resistant related proteins.Realtime PCR was used to detect changes in related genes.Results:Human colon cancer resistant cell lines were successfully established and named as HCT-116/L-OHP with a drug resistance index of 12.6.The inhibition rates of HCT-116 and HCT-1 16/L-OHP cells treated with cur(5,10,20,30,40 ?mol· L-1)at different concentrations showed no significant difference.Compared with HCT-116,the cell cycle of HCT-116/L-OHP was blocked in G1 phase,and the cell cycle of HCT-116/L-OHP was blocked in S phase and G2 phase after treatment with cur.The apoptosis rate of HCT-116 cells was 6.21%,and that of HCT-116/L-OHP cells was 8.36%.The apoptosis rate of drug-resistant cells treated with low,medium and high concentrations of cur was 10.65%,14.55%and 18.82%,respectively,which was significantly higher than that of the untreated group(p<0.01).Compared with HCT-116 cell lines,the expression level of resected and repaired cross complementation gene 1(ERCC1)protein and gene in HCT-116/L-OHP cell lines was significantly increased(P<0.01).The expression of b-cell lymphoma-2(Bcl-2),glutathione-s-transferase-consciousness(GST-consciousness),multidrug resistance-associated protein(MRP),p-glycoprotein(p-gp)and apoptotic inhibitory gene(Survivin)also increased significantly(P<0.01).After the action of curcumin(10,20,30?mol·L-1)of different concentrations,the expression of ERCC1 decreased(P<0.01),and the expression of Bcl-2?GST-??MRP?P-gp?Survivin also decreased to different degrees(P<0.05).Conclusion:HCT-116/L-OHP cell lines have stable drug resistance.The DNA synthesis of drug-resistant cells was blocked,the proliferation was slowed,and the apoptosis was increased.Its drug resistance mechanism may be up-regulated with the expression of ERCC1,which leads to the up-regulation of Bcl-2?GST-??MRP?P-gp?Survivin and other related proteins,and enables tumor cells to acquire drug resistance.Curcumin can reverse the drug resistance of HCT-116/L-OHP,and the proliferation of drug-resistant cells treated with cur was blocked.Its mechanism may be to reduce the expression of ERCC1,down-regulating the expression of Bcl-2?GST-??MRP?P-gp?Survivin and other drug-resistant related genes and proteins,and increase the sensitivity of tumor to L-OHP,so as to reverse the drug resistance of tumor cells.