The Mechanism And Effects Of The Tetrandrine Derivatives HL-49 Combined With Three Chemotherapeutic Drugs On Proliferation And Apoptosis In Triple-negative Breast Cancer MDA-MB-231 Cells

Objective: To investigate the effects of tetrandrine derivative HL-49 combined with cisplatin(DDP)or adriamycin(ADM)or paclitaxel,(TAX)on the proliferation and apoptosis of triple-negative breast cancer MDA-MB-231 cells,and to explore the possible mechanism.Methods: MDA-MB-231 cells were treated with different concentrations of HL-49,DDP or ADM alone and HL-49 combined with DDP or ADM or TAX.The proliferation inhibitory rate of MDA-MB-231 cells was detected by MTT assay and the Q value was calculated.The colony forming ability of MDA-MB-231 cells was detected by plate cloning method.The apoptosis rate was detected by FCM method and the Q value was calculated.Semi-quantitative RT-PCR assay was uesed to detect mRNA of expression of Bloom syndrome helicase(BLM),Breast cancer susceptibility gene 1(BRCA1)and DNA repair key enzyme Rad51,tumor suppressor gene p53,apoptosis related gene caspase 3,caspase 8,Bax,Bcl-2.The protein expression levels of Bloom's symdrome helicase(BLM),breast cancer susceptibility protein 1(BRCA1),DNA repair key enzyme Rad51,tumor suppressor gene p53,apoptosis-related protein caspase 3,caspase 8,caspase 10,Bax,and Bcl-2 were detected by Western blotting.Results: The proliferation inhibitory rate of MDA-MB-231 cells in HL-49 combined with DPP or ADM group was significantly higher than that in HL-49,DPP or ADM alone group(P < 0.05);the Q values of HL-49(1 ?mol/L and 2 ?mol/L)combined with DPP(5?mol/L and ?mol/L 10 ?mol/L)were 2.04,1.88,1.87 and 1.83;while the Q values of HL49(1?mol/L and 2 ?mol/L)combined with ADM(2?mol/L and 5 ?mol/L)were 1.37,1.21,1.36 and 1.23;the Q values of HL49(1?mol/L and 2 ?mol/L)combined with TAX(2?mol/L and 5 ?mol/L)were 2.86 ?1.80 ? 1.82 and 1.59.The colony formation number of MDA-MB-231 cells in HL-49 combined with DPP or ADM group was significantly lower than that in HL-49,DPP or ADM alone group(P < 0 05).The apoptosis rate of MDA-MB-231 cells in HL-49 combined with DPP or ADM group was higher than that in HL-49,DPP or ADM alone group(P < 0 05);the Q values of HL-49 combined with DPP were 1.20,1.35,1.70 and 2.06;the Q values of HL-49 combined with ADM were 1.48,1.17,1.99 and 2.13;the Q values of HL-49 combined with TAX were 1.33,1.01,1.98,1.58.The above results suggested that HL-49 and DDP or ADM or TAX had synergistic effects.As compared with HL-49,DPP or ADM alone group,the expression levels of caspase 3,caspase 8,and Bax mRNAs and proteins in HL49 combined with DDP or ADM group were up-regulated,and the expression levels of Rad51,BRCA1,BLM,p53 and Bcl-2 mRNAs and proteins were down-regulated(P < 0.05).Conclusion: Tetrandrine derivative HL-49 combined with DDP or ADM has synergetic effect.The anti-tumor activity of the combination treatment is related to the induction of apoptosis in triple-negative breast cancer MDA-MB-231 cells.