Effects of Rapamycin on Cell Apoptosis in MCF-7 Human Breast Cancer Cells

Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remainsunclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypotheticallyvia apoptotic promotion, using MCF-7 breast cancer cells. Materials and
Methods: MCF-7 cells were platedat a density of 15105 cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations ofrapamycin while only adding DMEM medium with PEG for the control regiment and grown at 37oC, 5% CO2and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated andrapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrastmicroscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation.In addition, cytotoxicity testing was performed using normal mouse breast mammary pads.
Results: Our resultsclearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The IC50 value of rapamycin onthe MCF-7 cells was determined as 0.4μg/ml (p<0.05). Direct observation by inverted microscopy demonstratedthat the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage,vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycleshowed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phasepopulations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively.
Conclusions: This study demonstrated thatrapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphologicalchanges of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis inlate stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin asan anti-cancer agent.