Marine Cooperates With Chemotherapy Drug And Reverses Multidrug Resistance In Breast Cancer Drug-resistant Cell Line MCF-7/ADR Through Inhibiting PI3K/AKT Signal Pathway

Objective To explore the cooperative effect of matrine and chemotherapy drug and the bioactivities of matrine and PI3K/AKT signal pathway in the reversal of multidrug resistance in breast cancer cells. Methods MTT assay was used to measure the inhibitory rate of 0.05g/L matrine with or without chemotherapy drug of different concertrations on breast cancer cell line MCF-7 after 24 h and measure the inhibitory rate of matrine and MK2206 of differernt concentrations on breast cancer drug resistant cell line MCF-7/ADR after 24 h. In addition, real-time RT-PCR and Western blotting were used to evaluate the m RNA and protein expression levels of MDR1,MRP1,PTEN,AKT and p-AKT in MCF-7/ADR cells. Results MTT assay results showed the inhibitory of chemotherapy drugs were increased with the increasing concentration and the inhibitory of chemotherapy drug with matrine groups were reduced obviously when compared with chemotherapy drug without matrine groups( P<0.05). Real-time RT-PCR results showed the relative expression levels of MDR1 and MRP1 in the 0.6 and 1.2g/L matrine groups were reduced gradually comparing with the control group( MDR1 0.659±0.074 and MRP1 0.503±0.058 in the 0.6 g/L matrine group vs MDR1 0.438±0.034 and MRP1 0.399±0.025 in the 1.2g/L matrine group, P < 0.01).The relative gene expression level of AKT was also reduced gradually,but that of PTEN was increased gradually. The expression levels of MDR1, MRP1 and AKT in the cells intervened separately by 0.6g/L matrine and 0.05umol/L, MK2206 didn’t have significant difference at the same inhibition rate. Western blotting results showed the expression levels of p-gp and MRP1 of the 0.3, 0.6 and 1.2g/L matrine groups were reduced gradually comparing with the control group,( p-gp 0.316±0.033 and MRP1 0.134±0.014 in the 0.6g/L matrine group, P < 0.01).The protein expression of p-AKT was also reduced gradually,but that of PTEN was increased gradually.The total AKT protein expression didn’t changed obviously(P>0.05). At the same inhibitory rate of matrine(0.6g/L) and MK2206(0.05umol/L),the marine group was more effective than MK2206 group in reducing the protein expression of p-gp,MRP1 and p-AKT and increasing the protein expression of PTEN. However,the total AKT protein expression of the two groups didn,t have significant difference(P>0.05).Conclusion Matrine has cooperative effect with chemotherapy and the effect of matrine mediating the reversal of drug resistance of breast cancer cells probably involves PI3K/AKT signal pathway.