Synergistic antitumor effect and reversal mechanism of K-selenocarrageenan in combination with adriamycin on HepG-2/ADR cells
Objective To explore the synergistic antitumor effect and reversal mechanism of κ-selenocarrageenan(KSC)in combination with adriamycin(ADR)on hepatoma resistant cell line HepG-2/ADR.Methods The multidrug resistance of HepG-2/ADR resistant cells to adriamycin(ADR),cisplatin(DDP)and paclitaxel(TAX)was detected by MTT assay,in order to evaluate the reversal effect of KSC on HepG-2/ADR cells.The effects of KSC and ADR on the cell cycle and apoptosis of HepG-2/ADR cells were detected by flow cytometry.Western blot was used to detect the expressions of drug-resistant proteins,cyclins and apoptosis-related proteins in HepG-2/ADR cells exposed to KSC and ADR.Moreover,the expressions of drug resistance related genes in HepG-2/ADR cells were further confirmed by RT-qPCR.Results MTT results showed HepG-2/ADR cells were resistant to ADR,TAX and DDP,demonstrating multi-drug resistance.KSC could reverse the multidrug resistance of HepG-2/ADR cells,and exhibited an additive effect while in combined with ADR on HepG-2/ADR cells.Flow cytometry indicated both KSC and ADR could induce apoptosis and S phase arrest of HepG-2/ADR cells.Additionally,western blot analysis showed KSC and ADR significantly downregulated the expressions of multidrug-resistant protein P-gp,MRP1 and ABCG2,as well as Cyclin A,Cyclin E,CDK2,Survivin and anti-apoptotic protein Bcl-2(P＜0.05 or P＜0.01),and significantly upregulated the expressions of pro-apoptotic proteins such as Bax,Caspase-3,Caspase-9,Cleaved-Caspase-3,and Cleaved-Caspase-9(P＜0.05 or P＜0.01).Besides,the combined group could synergistically promote or inhibit the expressions of apoptosis and cell cycle-related proteins.Furthermore,RT-qPCR demonstrated KSC and ADR alone or in combination could significantly downregulate the mRNA expression of P-gp,MRP1 and ABCG2.Conclusion KSC could effectively reverse multidrug resistance of hepatoma and synergistically enhance the sensitivity of HepG-2/ADR cells to adriamycin.The mechanism may be related to the induction of apoptosis and cell cycle arrest,as well as the downregulation of P-gp,MRP1 and ABCG2 expressions,thereby reversing the multidrug resistance in liver cancer.
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