Publication Date
Spring 5-10-2026
Keywords
Ewing Sarcoma, Pediatric Cancer, Everolimus, mTOR, Apoptosis, Proliferation, Migration, qRTPCR, Incucyte, Tissue Culture
Description
Ewing Sarcoma (ES) is an aggressive pediatric bone cancer characterized by rapid cell proliferation and poor prognosis, making the identification of therapeutic targets critical. One of the mechanistic targets is rapamycin (mTOR) signaling pathway, which is critical for regulating cell growth, proliferation, and survival. Abnormal activation of the mTOR signaling pathway has been linked to the progression of ES, as it drives uncontrolled cell growth and apoptosis resistance. Because mTOR represents a promising therapeutic target for ES treatment, we hypothesized that inhibiting mTOR activity through an siRNA-mediated knockdown or through Everolimus drug treatment, would reduce cell proliferation and promote ES cell apoptosis. Everolimus is an FDA-approved mTOR inhibitor that functions by binding to mTORC1 and blocking downstream signaling involved in cell growth and survival. To test this hypothesis, ES cells were treated with siRNA-mediated knockdown of mTOR and Everolimus at concentrations of 0.1µM and 1.0µM, and cell proliferation, apoptosis, and gene expression were measured using Incucyte live-cell imaging, Caspase 3/7 fluorescence, and qRT-PCR, respectively. Our results showed that mTOR inhibition successfully reduced mTOR gene expression and increased Caspase 3/7 activity, showing an increase in apoptosis, with the greatest effect observed at 1.0µM Everolimus together with siMTOR knockdown. Our findings support the hypothesis that mTOR inhibition reduces cell proliferation and promotes apoptosis in ES cells, suggesting that it is a potential therapeutic treatment for Ewing Sarcoma.
Collection
Cell and Molecular Methods
Format
Size or Duration
9 Pages
City
San Antonio
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Recommended Citation
Calvert, Dylan; Martinez, Malorie; Miner, Alexa; Quiroga, Ellis; and Shackleford, Terry Jo, "Everolimus and siRNA-Mediated mTOR Inhibition Reduces Proliferation and Promotes Apoptosis in Ewing Sarcoma Cells" (2026). Cell and Molecular Methods. 12.
https://commons.stmarytx.edu/biostulab25/12
Included in
Biology Commons, Cancer Biology Commons, Cell Biology Commons, Research Methods in Life Sciences Commons