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Terameprocol (Tetra-O-Methyl Nordihydroguaiaretic Acid), an Inhibitor of Sp1-Mediated Survivin Transcription, Induces Radiosensitization in Non-small.

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Presentation on theme: "Terameprocol (Tetra-O-Methyl Nordihydroguaiaretic Acid), an Inhibitor of Sp1-Mediated Survivin Transcription, Induces Radiosensitization in Non-small."— Presentation transcript:

1 Terameprocol (Tetra-O-Methyl Nordihydroguaiaretic Acid), an Inhibitor of Sp1-Mediated Survivin Transcription, Induces Radiosensitization in Non-small Cell Lung Carcinoma  Yunguang Sun, PhD, Nicholas J. Giacalone, BS, Bo Lu, MD, PhD  Journal of Thoracic Oncology  Volume 6, Issue 1, Pages 8-14 (January 2011) DOI: /JTO.0b013e3181fa646a Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

2 FIGURE 1 Terameprocol down-regulates survivin transciption and protein expression in HCC2429 and H460 cells. A, HCC2429 and H460 cells were transfected with 200 ng pLuc2931 or pLuc control and 0.6 μL lipofectamine Luciferase activity was detected 24 and 48 hours posttransfection and plotted. The luciferase assay was performed in triplicate, and error bar is shown as mean ± standard deviation (SD). B, Immunoblotting was used to determine survivin expression in HCC2429 and H460 cell lysates 24 and 48 hours after 0, 10, or 30 μM terameprocol treatment. Actin was probed to demonstrate equal loading. Journal of Thoracic Oncology 2011 6, 8-14DOI: ( /JTO.0b013e3181fa646a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

3 FIGURE 2 Radiation induces increased cleaved caspase-3 in HCC2429 cells but not H460 cells. A, HCC24249 and H460 cells were irradiated with 3 Gy. The expression of survivin, total caspase-3, and cleaved caspase-3 in HCC2429 and H460 cell lysates was determined 0, 24, 48, and 72 hours after radiation treatment by immunoblotting. Actin was probed to demonstrate equal loading. B, Immunoblotting was used to determine the expression of Bcl-2, Bcl-XL, Mcl-1, Bax, Bak, cIAP-1, cIAP-2, XIAP, and survivin in HCC2429 and H460 cell lysates. Tubulin was probed to demonstrate equal loading. Journal of Thoracic Oncology 2011 6, 8-14DOI: ( /JTO.0b013e3181fa646a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

4 FIGURE 3 Terameprocol induces increased radiosensitivity in HCC2429 and H460 cells but enhances apoptosis in only HCC2429 cells when used with radiation. A, HCC2429 and H460 cells were treated with 10 μM terameprocol or dimethyl sulfoxide (DMSO) for 24 hours, followed by irradiation with 0 to 6 Gy at 1.8 Gy/min. Forty-eight hours after irradiation, cells were cultured in drug-free media and incubated for 7 to 8 days. At this point, the surviving fraction and the drug enhancement ratio (DER) were determined. Each experiment was performed in triplicate, and data are shown as mean ± standard deviation (SD). B, HCC2429 and H460 cells were treated with 10 μM terameprocol or DMSO for 24 hours followed by radiation treatment with 0 or 3 Gy. Expression of survivin, total caspase-3, and cleaved caspase-3 was determined by immunoblotting 48 hours postradiation treatment. Actin was probed to demonstrate equal loading. Journal of Thoracic Oncology 2011 6, 8-14DOI: ( /JTO.0b013e3181fa646a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

5 FIGURE 4 Terameprocol treatment does not induce cell cycle changes in HCC2429 or H460 cells. HCC2429 and H460 cells were cultured with dimethyl sulfoxide (DMSO), 10 μM terameprocol, or 30 μM terameprocol for 24 or 48 hours, followed by ethanol fixation and staining with propidium iodide. Flow cytometry was used to determine percentage of HCC2429 or H460 cells in G1, S, or G2 phases of the cell cycle. The analysis was repeated three times using flow cytometry and the data graphed as a function of mean ± standard deviation (SD). Journal of Thoracic Oncology 2011 6, 8-14DOI: ( /JTO.0b013e3181fa646a) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

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