新型微管抑制剂Yf9b能够诱导Hela细胞G2/M期阻滞和凋亡Yf9b,a novel microtubule inhibitor,induced G2/M phase arrest and apoptosis in Hela cells
徐静雯,吴岑,沈继伟,张慧娟,左代英,李增强
XU Jing-wen,WU Cen,SHEN Ji-wei,ZHANG Hui-juan,ZUO Dai-ying,LI Zeng-qiang
摘要(Abstract)：
目的考察新型微管抑制剂2-甲氧基-5-(4-(3,4,5-三家氧基苯基)-1,3-硒唑-5-基)苯胺[2-methoxy-5-(4-(3,4,5-trimethoxyphenyl)-1,3-selenazol-5-yl)aniline,Yf9b]对多种人源性肿瘤细胞的抗肿瘤活性,并研究Yf9b抑制人宫颈癌Hela细胞增殖的机制。方法采用MTT法考察Yf9b对多种肿瘤细胞的增殖抑制作用。免疫荧光染色观察Yf9b与康普瑞丁(combretastatin A-4,CA-4)对微管的抑制作用。流式细胞技术考察Yf9b与CA-4对Hela细胞周期的影响,并考察Yf9b诱导Hela细胞凋亡的情况。结果 Yf9b的作用时间与剂量依赖性地抑制Hela细胞增殖,半数抑制浓度(IC50)为(42.9±2.7)nmol·L-1。Yf9b同母体化合物CA-4一样能抑制微管聚合,将Hela细胞阻滞在G2/M期,并最终诱导细胞凋亡。结论 Yf9b是一种新型微管抑制剂,能够诱导Hela细胞G2/M期阻滞和凋亡。
Objective In this study,we intended to investigate the antitumor activity of Yf9 b on different human cancer cell lines,and to study the mechanism of its cytotoxic upon Hela cells.Methods MTT assay was used to detect the cytotoxic effect of Yf9 b on different human cancer cell lines.Immunofluorescence staining was used to observe the change of microtubule in Hela cells after treated with Yf9 b and combretastatin(CA-4).Flowcytometry was applied to observe the change of cell cycle and apoptosis rate in Yf9 b or CA-4 treated Hela cells.Results MTT assay showed that Yf9 b exhibited time-and dose-dependent cell-killing effect on Hela cells,and the IC50 value of Yf9 b for Hela cells was(42.9 ± 2.7) nmol·L~(-1).Cell cycle analysis revealed that Yf9 b treatment resulted in cell cycle arrest at the G2 / Mphase in a timedependentmanner with subsequent apoptosis induction.Conclusions Yf9 b exhibits antitumor activity via the mechanism of disrupting the microtubule assembly,causingcell cycle arrest and consequently inducing apoptosis in Hela cells.
关键词(KeyWords)：
combretastatin A-4(CA-4);微管;凋亡;周期阻滞
combretastatin(CA-4);microtubule;apoptosis;cell cycle arrest
基金项目(Foundation):
作者(Author):
徐静雯,吴岑,沈继伟,张慧娟,左代英,李增强
XU Jing-wen,WU Cen,SHEN Ji-wei,ZHANG Hui-juan,ZUO Dai-ying,LI Zeng-qiang
DOI: 10.14066/j.cnki.cn21-1349/r.2017.01.013
参考文献(References)：
- [1]VALIRON O,CAUDRON N,JOB D.Microtubule dynamics[J].Cellular and Molecular Life Sciences,2001,58:2069-2084.
- [2]GARDNER M K,ZANIC M,HOWARD J.Microtubule catastrophe and rescue[J].Current Opinion in Cell Biology,2013,25:14-22.
- [3]FIELD J J,KANAKKANTHARA A,MILLER J H.Microtubule-targeting agents are clinically successful due to both mitotic and interphase impairment of microtubule function[J].Bioorganic&Medicinal Chemistry,2014,22:5050-5059.
- [4]EIKESDAL H P,BJERKVIG R,MELLA O,et al.Combretastatin A-4 and hyperthermia;a potent combination for the treatment of solid tumors[J].Radiotherapy and Oncology:Journal of the European Society for Therapeutic Radiology and Oncology,2001,60:147-154.
- [5]ZWEIFEL M.JAYSON G C,REED N S,et al.Phase II trial of combretastatin A4 phosphate,carboplatin,and paclitaxel in patients with platinum-resistant ovarian cancer[J].Annals of Oncology:Official Journal of the European Society for Medical Oncology/ESMO,2011,22:2036-2041.
- [6]KAPOOR S.Combretastatin A4 and its emerging antineoplastic effects[J].Surgery,2013,153:881.
- [7]WALLENBERG M,MISRA S,WASIK A M,et al.Selenium induces a multi-targeted cell death process in addition to ROS formation[J].Journal of Cellular and Molecular Medicine,2014,18:671-684.
- [8]PARK S O,YOO Y B,KIM Y H,et al.Effects of combination therapy of docetaxel with selenium on the human breast cancer cell lines MDA-MB-231 and MCF-7[J].Annals of Surgical Treatment and Research,2015,88:55-62.
- [9]ACHARYA B R,BHATTACHARYYA S,CHOUDHURY D,et al.The microtubule depolymerizing agent naphthazarin induces both apoptosis and autophagy in A549 lung cancer cells[J].Apoptosis,2011,16:924-939.
- [10]ROGALSKA A,SZULA E,GAJEK A,et al.Activation of apoptotic pathway in normal,cancer ovarian cells by epothilone B[J].Environmental Toxicology and Pharmacology,2013,36:600-610.
- [11]CHUMDURI C,GILLISSEN B,RICHTER A,et al.Apoptosis resistance,mitotic catastrophe,and loss of ploidy control in Burkitt lymphoma[J].Journal of Molecular Medicine,2015,93:559-572.
- [12]QIAO F,ZUO D,SHEN X,et al.DAT-230,a novel microtubule inhibitor,exhibits potent anti-tumor activity by inducing G2/M phase arrest,apoptosis in vitro and perfusion decrease in vivo to HT-1080[J].Cancer Chemother Pharmacol,2012,70:259-270.
- [13]ZUO D,GUO D,JIANG X,et al.3-(3-Hydroxy-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)-1,2,5-selenadiazole(G-1103),a novel combretastatin A-4 analog,induces G2/M arrest and apoptosis by disrupting tubulin polymerization in human cervical Hela cells and fibrosarcoma HT-1080 cells[J].Chemico-biological Interactions,2015,227:7-17.