Wolmarans Elize, Mqoco Thandi V, Stander Andre, Nkandeu Sandra D, Sippel Katherine, McKenna Robert, Joubert Annie
Department of Physiology, University of Pretoria, Pretoria, South Africa.
Cell Mol Biol Lett. 2014 Mar;19(1):98-115. doi: 10.2478/s11658-014-0183-7. Epub 2014 Feb 22.
Cancer is the second leading cause of death in South Africa. The critical role that microtubules play in cell division makes them an ideal target for the development of chemotherapeutic drugs that prevent the hyperproliferation of cancer cells. The new in silico-designed estradiol analogue 2-ethyl-3-O-sulfamoylestra-1,3,5(10)16-tetraene (ESE-16) was investigated in terms of its in vitro antiproliferative effects on the esophageal carcinoma SNO cell line at a concentration of 0.18 μM and an exposure time of 24 h. Polarization-optical differential interference contrast and triple fluorescent staining (propidium iodide, Hoechst 33342 and acridine orange) revealed a decrease in cell density, metaphase arrest, and the occurrence of apoptotic bodies in the ESE-16-treated cells when compared to relevant controls. Treated cells also showed an increase in the presence of acidic vacuoles and lysosomes, suggesting the occurrence of autophagic processes. Cell death via autophagy was confirmed using the Cyto-ID autophagy detection kit and the aggresome detection assay. Results showed an increase in autophagic vacuole and aggresome formation in ESE-16 treated cells, confirming the induction of cell death via autophagy. Cell cycle progression demonstrated an increase in the sub-G1 fraction (indicative of the presence of apoptosis). In addition, a reduction in mitochondrial membrane potential was also observed, which suggests the involvement of apoptotic cell death induced by ESE-16 via the intrinsic apoptotic pathway. In this study, it was demonstrated that ESE-16 induces cell death via both autophagy and apoptosis in esophageal carcinoma cells. This study paves the way for future investigation into the role of ESE-16 in ex vivo and in vivo studies as a possible anticancer agent.
癌症是南非第二大死因。微管在细胞分裂中发挥的关键作用使其成为开发预防癌细胞过度增殖的化疗药物的理想靶点。对新的计算机辅助设计的雌二醇类似物2-乙基-3-O-氨磺酰基雌甾-1,3,5(10),16-四烯(ESE-16),在浓度为0.18 μM、暴露时间为24小时的条件下,对食管癌细胞系SNO进行了体外抗增殖作用研究。偏振光微分干涉对比和三重荧光染色(碘化丙啶、Hoechst 33342和吖啶橙)显示,与相关对照相比,ESE-16处理的细胞中细胞密度降低、中期停滞以及凋亡小体出现。处理后的细胞中酸性液泡和溶酶体的数量也有所增加,表明存在自噬过程。使用Cyto-ID自噬检测试剂盒和聚集体检测试验证实了通过自噬导致的细胞死亡。结果显示ESE-16处理的细胞中自噬液泡和聚集体形成增加,证实了通过自噬诱导细胞死亡。细胞周期进程显示亚G1期部分增加(表明存在凋亡)。此外,还观察到线粒体膜电位降低,这表明ESE-16通过内源性凋亡途径诱导凋亡性细胞死亡。在本研究中,证明了ESE-16在食管癌细胞中通过自噬和凋亡诱导细胞死亡。本研究为未来研究ESE-16作为一种可能的抗癌剂在离体和体内研究中的作用铺平了道路。