Tangeretin derivative, 5-acetyloxy-6,7,8,4-tetramethoxyflavone induces G2/M arrest, apoptosis and autophagy in human non-small cell lung cancer cells in vitro and in vivo
Tangeretin derivative, 5-acetyloxy-6,7,8,4-tetramethoxyflavone induces G2/M arrest, apoptosis and autophagy in human non-small cell lung cancer cells in vitro and in vivo
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DOI：
10.1080/15384047.2015.1108491
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发表时间：
2016-01-01
影响因子：
3.6
通讯作者：
Lin, Chi-Chen
中科院分区：
文献类型：
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作者：
Li, Yi Rong;Li, Shiming;Lin, Chi-Chen
橘皮素是一种重要的中草药橘皮中的主要植物化学物质，已被发现具有抗癌特性。为了提高橘子素的生物利用度和效力，合成了其衍生物 5-乙酰氧基-6,7,8,4-四甲氧基黄酮 (5-AcTMF)，并显示出对人乳腺癌和白血病癌细胞系的增殖活性的有效抑制作用。在本研究中，我们进一步研究了 5-AcTMF 在体外和体内对 CL1-5 非小细胞肺癌细胞 (NSCLC) 的抗癌作用，并证明 5-AcTMF 有效抑制癌细胞增殖，诱导与 cdc2 和 CDC25c 相关的 G2/M 期阻滞，并增加与 caspase 激活相关的凋亡细胞，下调 Bcl-2、XIAP Survivn，诱导细胞色素 c 释放到细胞质中并破坏线粒体膜电位。我们还发现 CL1-5 的 5-AcTMF 处理激活了自噬，表现为触发自噬体形成、LC3-II 水平增加和 LC3 点的形成。此外，我们还发现5-AcTMF降低了磷脂酰肌醇3-激酶/AKT/mTOR信号通路。通过 AKT cDNA 转染过度表达 AKT 可减少 5-AcTMF 介导的细胞凋亡和自噬，支持通过抑制 AKT 途径诱导细胞凋亡和自噬。在一项动物研究中，5-AcTMF 有效延迟了 CL1-5 异种移植裸鼠模型中的肿瘤生长，且未观察到不良反应。免疫组织化学分析表明，5-AcTMF 在体内诱导 CL1-5 细胞凋亡和自噬。综上所述，这些数据表明 5-AcTMF 是一种新型小分子药物，可以抑制 NSCLC 细胞增殖，并通过线粒体凋亡途径和自噬诱导 G(2)/M 期阻滞。
Tangeretin, a major phytochemicals in tangerine peels - an important Chinese herb, has been found to have anti-carcinogenic properties. To improve bioavailability and increase potency of tangeretin, its derivative, 5-acetyloxy-6,7,8,4-tetramethoxyflavone (5-AcTMF), has been synthesized and shown potent inhibition of proliferation activity against human breast and leukemia cancer cell lines. In this study, we have further investigated the anticancer effects of 5-AcTMF on CL1-5 non-small cell lung cancer cells (NSCLC) both in vitro and in vivo and demonstrated that 5-AcTMF effectively inhibited cancer cell proliferation, induced G2/M-phase arrest associated with cdc2 and CDC25c and increased in the apoptotic cells associated with caspase activation, down regulation of Bcl-2, XIAP and Survivn, inducing release of cytochrome c into the cytosol and disruption of mitochondrial membrane potential. We also found that 5-AcTMF treatment of CL1-5 activated autophagy, indicated by triggered autophagosome formation and increased LC3-II levels and formation of LC3 puncta. Moreover, we also found that 5-AcTMF lowered phophoatidylinositol 3-kinase/AKT/mTOR signaling pathway. Over-expression of AKT by AKT cDNA transfection decreased 5-AcTMF mediated apoptosis and autophagy, supporting the induction of apoptosis and autophagy by inhibition of AKT pathway. In an animal study, 5-AcTMF effectively delayed tumor growth in a nude mouse model of CL1-5 xenografts without observed adverse effect. Immunohistochemistry Analysis indicated that 5-AcTMF induced CL1-5 cell apoptosis and autophagy in vivo. Taken together, these data demonstrate that 5-AcTMF is a novel small molecule agent that can inhibit NSCLC cell proliferation, and induce G(2)/M phase arrest and via the mitochondrial apoptotic pathway and autophagy.