Reactive oxygen species and senescence modulatory effects of rice bran extract on 4T1 and NIH-3T3 cells co-treatment with doxorubicin

Objective:To determine the effect of rice bran extract(RBE)in combination with doxorubicin on 4 T1 triple-negative breast cancer cells and NIH-3 T3 cells.Methods:RBE was obtained by maceration with n-hexane.The phytochemical profile of RBE was observed using highperformance liquid chromatography.Cytotoxic effect of RBE was evaluated through MTT assay.In addition,flow cytometry was used for cell cycle and apoptosis analysis.Cellular senescence was observed using SA-β-Gal assay and intracellular reactive oxygen species(ROS)levels were evaluated using DCFDA staining.The pro-oxidant property of RBE was also evaluated through 1-chloro-2,4-dinitrobenzene spectrophotometry and molecular docking.Results:RBE was obtained with a yield of 18.42%w/w and contained tocotrienols as the major compound.RBE exerted no cytotoxic effect on 4 T1 and NIH-3 T3 cells.However,RBE in combination with doxorubicin decreased 4 T1 cell viability synergistically(combination index<0.9)and induced apoptosis and senescence on 4 T1 cells.RBE significantly decreased senescence in doxorubicin-treated NIH-3 T3 cells.Additionally,RBE did not increase ROS levels in doxorubicin-treated 4 T1 cells.Meanwhile,the combination of RBE and doxorubicin reduced ROS levels in NIH-3 T3 cells.RBE significantly reduced glutathione-S-transferase activity and alpha-tocotrienol interacted with glutathione-Stransferase in the glutathione binding site.Conclusions:Rice bran may be used as a co-chemotherapeutic agent to improve the therapeutic effectiveness of doxorubicin while protecting against the cellular senescence effects of doxorubicin on healthy cells.