Cell based target inhibition
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HT-1080 cells are treated with increasing concentrations of Deforolimus (0-100 nM) for 2 hours, prior to harvest. Cellular lysates are extracted in denaturing lysis buffer, resolved on SDS-PAGE and transferred to PVDF membranes. After blocking, membranes are incubated with primary antibodies for 1 hour, followed by appropriate HRP-conjugated secondary antibodies for 1 hour at room temperature. Immunoreactive proteins are detected using enhanced chemiluminescence and autoradiography performed by exposure to X-ray film. IC50 is determined from the inhibition of levels of phosphorylated ribosomal protein S6 (p-S6) and 4E-BP1 (p-4E-BP1).
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Cell lines
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HCT-116, SK-UT-1, HT-1080, SW872, MCF7, SK-LMS-1, U-87, A-204, PC-3, Endothelial cells, SK-UT-1B, ARK1 and ARK2 cells
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Applications
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Ridaforolimus showed the broad inhibitory effects on cell growth, division, metabolism, and angiogenesis and attenuated mTOR signaling [1]. Moreover, Ridaforolimus (20–100 nM) treatment decreased the viability in ARK1 and ARK2 cells [2].
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Applications
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Ridaforolimus induced tumor growth inhibition in mice bearing PC-3 (prostate), HCT-116 (colon), MCF7 (breast), PANC-1 (pancreas), SK-LMS-1 (sarcoma) or A549 (lung) xenografts [1]. Moreover, Ridaforolimus improved the anti-tumor activity of dual HER2 blockade in mice harboring uterine serous carcinoma (USC) xenografts [2].
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References:
1. Rivera, V. M., Squillace, R. M., Miller, D., Berk, L., Wardwell, S. D., Ning, Y., Pollock, R., Narasimhan, N. I., Iuliucci, J. D., Wang, F. and Clackson, T. (2011) Ridaforolimus (AP23573; MK-8669), a potent mTOR inhibitor, has broad antitumor activity and can be optimally administered using intermittent dosing regimens. Mol Cancer Ther. 10, 1059-1071
2. Hernandez, S. F., Chisholm, S., Borger, D., Foster, R., Rueda, B. R. and Growdon, W. B. (2016) Ridaforolimus improves the anti-tumor activity of dual HER2 blockade in uterine serous carcinoma in vivo models with HER2 gene amplification and PIK3CA mutation. Gynecol Oncol. 141, 570-579
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