Synergistic Effect and Mechanism of Shugan Huatan Sanjie Recipe and Paclitaxel Against Breast Cancer MCF-7/PTX Cells
Objective To investigate the synergistic effect of Shugan Huatan Sanjie prescription(SHSF)and paclitaxel on MCF-7/PTX cells of breast cancer and its mechanism.Methods The PTX-resistant breast cancer cell line MCF-7/PTX was established by continuous induction of low concentration.The effects of different concentrations of PTX on MCF-7 and MCF-7/PTX cells.Then the proliferation of MCF-7/PTX cells by SHSF containing serum were detected by MTT assay,and the drug resistance index(RI)and reversion times were calculated according to the IC50 value.MCF-7/PTX cells were divided into blank control group,PTX group(45 nmol·L-1 PTX),SHSF group(7.5％SHSF drug-containing serum)and PTX+SHSF group(45 nmol·L-1 PTX+7.5％SHSF drug-containing serum).The cells were treated with drugs for 24 h.The apoptosis level of each group was detected by flow cytometry.The expression levels of apoptotic proteins Bax,Bcl-2,p-Akt(ser473),Akt,p-mTOR(Ser2448)and mTOR were detected by Western blot.Results The PTX-resistant cell line MCF-7/PTX was established successfully,and the RI value was 6.70.The proliferative activity of MCF-7/PTX cells decreased in a concentration-dependent manner with the increase of SHSF drug-containing serum concentration.And the resistance reversal ratio of 7.5％SHSF serum to MCF-7/PTX cells was 3.48.Compared with blank control group,the apoptosis levels of MCF-7/PTX cells in PTX group and SHSF group were significantly increased(P＜0.01),the protein expression level of Bax was significantly up-regulated(P＜0.01),and the protein expression levels of Bcl-2,p-Akt/Akt and p-mTOR/mTOR were significantly down-regulated(P＜0.05);Compared with PTX group,the apoptosis level in PTX+SHSF group was significantly increased(P＜0.01),the expression level of Bax protein was significantly up-regulated(P＜0.01),and the expression levels of Bcl-2,p-Akt and p-mTOR protein were significantly down-regulated(P＜0.01).Conclusion Shugan Huatan Sanjie Recipe promotes apoptosis by inhibiting Akt/mTOR pathway,and thus plays a synergistic effect with paclitaxel on MCF-7/PTX cells of breast cancer.
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