Investigation On The Antiproliferation Effect And Its Mechanism Of Cardamonin Combined With Cisplatin On SKOV3Cells

Background and ObjectivePlatinum-based chemotherapy is an important method in the treatment ofovarian cancer. However, the efficacy of these chemotherapy drugs is severelyrestricted by the development of tumor chemoresistance in chemotherapy. Currently,use other antineoplastic drugs in combination with platinum drugs to achieveefficacy is commonly treatment in clinical. Therefore, searching for chemotherapydrugs that can improve the efficacy of platinum drugs becomes key factor in thetreatment of ovarian cancer. Cardamonin (CAR) is a flavonoid, which is isolatedfrom the seed of Alpiniakatsumadai. It has been demonstrated that CAR possessesdiverse pharmacologic effects, such as anti-tumor and anti-inflammatory. CARpotentiates the anti-tumor effect of TRAIL, which mediated through up-regulation ofdeath receptors (DRs) and apoptosis proteins. However, it is still unclear if CARpotentiates the anti-tumor effect of cisplatin (Cis). In the present study, weinvestigate the effect of CAR combined with Cis on the proliferation of ovariancancer cell line SKOV3, we also evaluate the effectiveness of CAR to potentiate theanti-tumor effect of Cis and explore the mechanism, thus these allow us to show newdirections to develop anti-tumor drugs used for combination with platinum-basedchemotherapy, and provide experimental evidence and theoretical foundation for thefurther development of CAR.Methods1. Groups: Control group, solvent control group (0.1%DMSO), Cis group (2μg/mL), CAR group (20μM), and Cis+CAR group (2μg/mL+20μM) were setin the SKOV3cells.2. Measurements and methodology:2.1Cell proliferation was assessed by MTT method (λ=490nm);2.2The cell cycle was detected by PI fluorescent staining method;2.3The autophagy level of cells was detected by MDC staining method;2.4Expressions of mRNA encoding glutathione S-transferase-π (GST-π) andmultidrug resistance protein (MRP) were assayed by Real-time PCR method;2.5The expressions of Bcl-2, XIAP, survivin and LC3were determined byWestern-blotting analysis.Results 1. The proliferation of SKOV3cells was markedly inhibited by CAR incombination with Cis, the inhibition rate is higher than Cis group significantly(P <0.01), and there were synergistic effects between CAR (20μM) and Cis (2μg/mL).2. The colony formation of SKOV3cells was markedly inhibited by CARcombined with Cis, and the inhibition rate is higher than Cis group significantly(P<0.01).3. Compared with single drug group, combination group did not make significantdifference on the generation of DNA fragments.4. Cis could induce expression of GST-π mRNA(P <0.01), while it had nosignificant influence on expression of MRP. CAR could down-regulateexpression of MRP mRNA, while it had no significant influence on expressionof GST-π.5. The expression of Bcl-2, XIAP and survivin were decreased by both Cis andCAR (P <0.01); while expression of Bcl-2, XIAP and survivin weresignificantly decreased in combination group, and the inhibition effect washigher than single drug group significantly.6. The fluorescent spots of MDC staining increased in cells treated by Cis or CARrespectively; while the stained spots significantly elevated in cells treated withCAR and Cis, thus the combination of Cis and CAR induced autophagysynergistically.Conclusion1. CAR and Cis inhibit proliferation of SKOV3cells synergistically.2. CAR combined with Cis enhance autophagy, and inhibit the cell cycle of SKOV3cells.3. The synergistic effect of CAR to enhance anti-tumor effect of Cis is associatedwith down-regulation of anti-apoptotic protein and drug resistance gene closely.