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Citronellol Induces the Necroptosis of Human Lung Cancer Cell via TNF-α pathway Through RIP3 Activation and Reactive Oxygen Species Accumulation
|關鍵字:||香茅醇;非小細胞肺癌;程序性細胞壞死;活性氧自由基;Citronellol;Necroptosis;Human Lung Cancer Cell;TNF-α;RIP3;ROS||引用:||1. 衛生福利部. (2014). 2. Travis, W.D., Travis, L.B. & Devesa, S.S. Lung cancer. Cancer 75, 191-202 (1995). 3. Travis, W.D. Pathology of lung cancer. Clin Chest Med 23, 65-81, viii (2002). 4. Subramanian, J. & Govindan, R. Lung cancer in never smokers: a review. J Clin Oncol 25, 561-70 (2007). 5. Raz, D.J., He, B., Rosell, R. & Jablons, D.M. Bronchioloalveolar carcinoma: a review. Clin Lung Cancer 7, 313-22 (2006). 6. Vaporciyan, A.N.J., Lee JS Cancer Medicine. (2000). 7. Grand, B. et al. High grade neuroendocrine lung tumors: pathological characteristics, surgical management and prognostic implications. Lung Cancer 81, 404-9 (2013). 8. Plus, M. 9. Barbone, F.B.M., Cavallieri F, Stanta G. Cigarette smoking and histologic type of lung cancer in men. American College of Chest Physicians (1997). 10. Collins, L.G., Haines, C., Perkel, R. & Enck, R.E. Lung cancer: diagnosis and management. 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Our previous studies showed that the essential oils of Pelargonium capitatum (EOPC) could induce the necroptosis of human non-small lung cancer cells. Citronellol, the major compound in EOPC, has anti-bacterial, anti-oxidant and anti- inflammatory effects. In this research, we would like to prove that citronellol may have noteworthy anticancer activity parallel to that of EOPC.
Citronellol was used to treat lung cancers (A549, H1299, H23), breast cancer (BT-20), prostate cancer (PC3) and breast normal cell (MCF10A) to measure the cytotoxicity of citronellol with MTT assay. Among the six cell lines, human non-small lung cancer H1299 was shown the most susceptibility to citronellol (24/48/72 hr IC50=48 μg/mL, 39 μg/mL, 26 μg/mL). PI staining experiment with flow cytometry were applied to citronellol-treated H1299 cells.We found that citronellol could inhibit the cell growth and arrest the cell cycle at G1 phase. Further, citronellol could induce the necroptosis of H1299 cells by PI/Annexin-V double staining experiment. Due to necroptosis was resulted from ROS accumulation, H2DCF-DA fluorescent staining was applied to detect ROS products in H1299 cells after treated cells with citronellol and found that ROS concentration was accumulated in cancer cellswith time-dependent manner.
The references indicated that when the necroptosis happened, TNF-α would be activated, down-stream RIP3 would complex with RIP1, and then ROS would accumulate quickly in cells. ROS accumulation caused DNA damage, followed by LMP and PARP over-activation andcellular energy depletion. Moreover, lysosome would be destroyed, and caused cathepsin B/D proteins to release into cytoplasm. If the necroptosis of signal pathway occurred, caspase would not be activated. Data of western blotting experiments showed that citronellol regulated the cell cycle of cancer cells by decreasing the protein levels of cyclin E, and promoted intracellular ROS accumulation. Citronellol could also increase the protein levels of TNF-α, RIP3, pro-PARP and cathepsin B/D in treated cells. However the protein levels of caspase-3 was not activated. We confirm that citronellol could induce the necroptosis of human lung cancer cell H1299 via TNF-α pathway through the RIP3 activation and ROS accumulation.
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