Tavakoli-Yaraki Masoumeh, Karami-Tehrani Fatemeh, Salimi Vahid, Sirati-Sabet Majid
Cancer Research Laboratory, Department of Clinical Biochemistry, School of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-331, Tehran, Iran.
Tumour Biol. 2013 Feb;34(1):241-9. doi: 10.1007/s13277-012-0544-7. Epub 2012 Oct 6.
15-Lipoxygenase-1 (15-Lox-1) is a key enzyme mediating oxidative metabolism of polyunsaturated fatty acids and has attracted considerable interest as a potential target for the induction of apoptosis in cancer cells. Knowledge of relationship between 15-Lox-1 and histone deacetylase inhibitors is lacking in the breast cancer. This study is aimed to investigate the role of Trichostatin A (TSA) and 13(S)-HODE, as a metabolite of 15-Lox-1, in the regulation of breast cancer cell growth. The cytotoxic effect of TSA, as a potent HDAC inhibitor, was measured using MTT assay. Annexin V-FITC and PI staining were performed to detect apoptosis and cell cycle distribution using Flow cytometry. The role of 15-Lox-1 in the regulation of cell growth was assessed by 15-Lox-1 inhibitor and the level of 15-Lox-1 metabolite was measured to determine 15-Lox activity after treatment by TSA. The results demonstrated that TSA induced cell growth inhibition via 15-Lox-1, in a dose- and time-dependent manner, and subsequently accompanied by the cell cycle arrest and induction of apoptosis. Moreover, growth inhibitory effect of TSA was associated with the elevation of 15-Lox-1 metabolite (13(S)-HODE). This study provided evidences that the inhibitory effect of TSA on the breast cancer cell growth occurs via the induction of 15-Lox-1 activity and 13(S)-HODE production. Our findings underline the possible role of 15-Lox-1/13(S)-HODE pathway as a promising molecular approach for the induction of apoptosis in breast cancer cells.
15-脂氧合酶-1（15-Lox-1）是介导多不饱和脂肪酸氧化代谢的关键酶，作为诱导癌细胞凋亡的潜在靶点已引起广泛关注。在乳腺癌中，关于15-Lox-1与组蛋白脱乙酰酶抑制剂之间关系的了解尚少。本研究旨在探讨曲古抑菌素A（TSA）和15-Lox-1的代谢产物13(S)-羟基十八碳二烯酸（13(S)-HODE）在调节乳腺癌细胞生长中的作用。使用MTT法检测TSA（一种有效的组蛋白脱乙酰酶抑制剂）的细胞毒性作用。采用膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）和碘化丙啶（PI）染色，通过流式细胞术检测细胞凋亡和细胞周期分布。用15-Lox-1抑制剂评估15-Lox-1在细胞生长调节中的作用，并检测15-Lox-1代谢产物的水平，以确定TSA处理后15-Lox的活性。结果表明，TSA通过15-Lox-1以剂量和时间依赖性方式诱导细胞生长抑制，随后伴随着细胞周期阻滞和凋亡诱导。此外，TSA的生长抑制作用与15-Lox-1代谢产物（13(S)-HODE）的升高有关。本研究提供的证据表明，TSA对乳腺癌细胞生长的抑制作用是通过诱导1,5-Lox-1活性和产生13(S)-HODE实现的。我们的研究结果强调了15-Lox-1/13(S)-HODE途径作为诱导乳腺癌细胞凋亡的一种有前景的分子方法的可能作用。
