A novel adamantane thiadiazole derivative induces mitochondria-mediated apoptosis in lung carcinoma cell line
A novel adamantane thiadiazole derivative induces mitochondria-mediated apoptosis in lung carcinoma cell line
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DOI：
10.1016/j.bmc.2016.10.040
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发表时间：
2017-01-01
影响因子：
3.5
通讯作者：
Mohamed, Mervat S.
中科院分区：
文献类型：
--
作者：
Ali, Ayat G.;Mohamed, Magda F.;Mohamed, Mervat S.
有机化合物与细胞凋亡调节蛋白的相互作用是一个有吸引力的研究领域，因为它与开发新的癌症化疗药物有关。我们设计了四种新的金刚烷噻二唑衍生物（ATDs）。通过分子建模，理论上测试了这四种ATDs与细胞凋亡抑制蛋白模型的结合亲和力。合成了结合亲和力最高的ATD-4，并对其进行了表征。测定了ATD-4对不同肿瘤细胞系和正常细胞系的体外细胞毒性，并与5-氟尿嘧啶作为标准阳性对照进行了比较。选择经ATD-4处理后细胞毒活性最高的肺癌细胞系，进一步研究ATD-4是否如分子模型所预期的那样通过诱导凋亡发挥细胞毒活性。观察ATD-4诱导肺癌细胞凋亡的各种生化和形态学特征。生化：流式细胞术检测ATD-4对细胞周期的影响及诱导凋亡的能力。用比色法检测Caspase-3活性。采用实时聚合酶链反应（Real timepolymerase chain reaction, q-PCR）检测p53、caspase-3、bcl-2和bax基因的表达。形态学：使用原子力显微镜（AFM）检测细胞表面形态、肉芽和平均表面粗糙度的变化。透射电镜观察细胞收缩、细胞器增多、线粒体数目及形态改变、染色质凝聚、膜泡及凋亡小体形成。结果表明，ATD-4对A549细胞的抗肿瘤活性是通过诱导线粒体凋亡途径发挥的。(C) 2016 Elsevier Ltd.版权所有。
The interaction of organic compounds with apoptosis regulatory proteins is an attractive field of research because of its relevance in the development of new chemotherapeutic agents for cancer treatment. Our group designed four new adamantane thiadiazole derivatives (ATDs). The four ATDs were theoretically tested for their binding affinities to a model of an apoptosis inhibitor protein using molecular modeling. ATD-4 which interacted with the highest binding affinity was synthesized and characterized. The in vitro cytotoxicity of ATD-4 against different cancer cell lines as well as normal cell line was determined and compared with 5-fluorouracil as a standard positive control. The lung carcinoma cell line that showed the highest cytotoxic activity due to ATD-4 treatment was chosen to further study if ATD-4 can perform its cytotoxic activity through the induction of apoptosis as expected from molecular modeling. Inducing apoptosis by ATD-4 in lung carcinoma cell line was assessed by various biochemical and morphological characteristics. Biochemically: The effect of ATD-4 on cell cycle and its ability to induce apoptosis were checked through flow cytometry. Caspase-3 activity was detected by a colorimetric method. Real timepolymerase chain reaction (q-PCR) was used to detect p53, caspase-3, bcl-2 and bax gene expression. Morphologically: Changes in cell surface morphology, granulation and average surface roughness were detected using atomic force microscopy (AFM). Cell shrinkage, increase in cytoplasmic organelles, changes in mitochondrial number and morphology, chromatin condensation, membrane blebbing and formation of apoptotic bodies were detected using transmission electron microscopy (TEM). The obtained results suggest that ATD-4 exerted its antitumor activity against A549 cells through the induction of the intrinsic (mitochondrial) apoptotic pathway. (C) 2016 Elsevier Ltd. All rights reserved.