Effect of MTOR Inhibitor Combined with Photodynamic Therapy on the Regulation of Cancer Stem Cells through Reactive Oxygen Species
Effect of MTOR Inhibitor Combined with Photodynamic Therapy on the Regulation of Cancer Stem Cells through Reactive Oxygen Species
Yakun Ge
ABSTRACT
To explore the mechanism of mammalian target of rapamycin (mTOR) inhibitor combined with photodynamic therapy to regulate cancer stem cells through reactive oxygen species (ROS). A2780 cell line was purchased from the American Type Culture Collection and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 50μunits/mL penicillin and 50μg/mL streptomycin, maintained at 37°C and 5% CO2 in a humid incubator environment. Quantitative analysis of ROS was performed using flow cytometry and median fluorescence intensity detection. Cell proliferation was detected by MTT method. Combined treatment promoted the production of ROS. The cell viability of the combined treatment group was lower than that of the control group at 24h, 48hs and 72h (P<0.05). Compared with the control group, the expression level of E-cadherin mRNA in the combined treatment group increased (P<0.05), and the expression levels of N-cadherin and vimentin decreased (P<0.05). Compared with the control group, the apoptosis rate of the combined treatment group increased (P<0.05). Compared with the control group, the expression levels of Bax, caspase-3 and caspase-9 were higher in the combined treatment group (P<0.05), and the expression levels of Bcl-2, PI3K, Akt and mTOR were lower (P<0.05). mTOR inhibitor combined with photodynamic therapy increased the generation of ROS to inhibit PI3K/Akt/mTOR signal transduction, increase the expression levels of pro-apoptotic proteins Bax, caspase-3 and caspase-9, inhibit the expression level of anti-apoptotic protein Bcl-2 and the epithelial-mesenchymal transition of cancer stem cells, reduce the proliferation ability of cancer stem cells, and increase the apoptosis of cancer stem cells.
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