Stellera chamaejasme against multidrug resistance of triple-negative breast cancer MDA-MB-231 cell through Nrf2
This study aims to investigate the effect and mechanism of Stellera chamaejasme extract(SCL)on multidrug resistance(MDR)in breast cancer.Human triple-negative breast cancer cell line MDA-MB-231 and its adriamycin-resistant cell line MDA-MB-231/ADR were used in the experiment.Cell viability was detected by methyl thiazolyl tetrazolium(MTT)assay,and cell apoptosis was detected by DAPI staining and Annexin-V/Pi double staining.Western blot(WB)was used to detect the expression levels of Keapl,Nrf2,HO-1,Bcl-2,Bax,caspase-9,and caspase-3.Immunofluorescence staining was used to observe the distribution of Nrf2 in the cell,and flow cytometry was used to detect the level of reactive oxygen species(ROS)in the cell.The results showed that the resis-tance factor of SCL was 0.69,and that of adriamycin and paclitaxel was 8.40 and 16.36,respectively.DAPI staining showed that SCL could cause nuclear shrinkage and fragmentation of breast cancer cells.Annexin-V/Pi double staining showed that the average apopto-sis rate of the drug-resistant cells was 32.64％and 50.29％,respectively under medium and high doses of SCL.WB results showed that SCL could significantly reduce the expression levels of anti-apoptotic proteins Bcl-2,caspase-9,and caspase-3 and significantly in-crease the expression level of pro-apoptotic protein Bax.Further studies showed that SCL could significantly promote the expression of Keap1,significantly inhibit the expression of Nrf2 and HO-1,and significantly reduce the expression level of Nrf2 in the nucleus.Cor-respondingly,flow cytometry showed that the intracellular ROS level was significantly increased.In conclusion,SCL can significantly inhibit the proliferation of MDA-MB-231 multidrug-resistant cells of triple-negative breast cancer and cause cell apoptosis,and the mechanism is related to inhibiting Keapl/Nrf2 signaling pathway,leading to ROS accumulation in drug-resistant cells and increasing the expression of apoptosis-related proteins.
Stellera chamaejasme extractmultidrug resistance in breast canceapoptosisNrf2antioxidant
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