摘要
背景:XPO1(exportin 1)是核质转运的重要介质之一,在多种人类恶性肿瘤中表达增高,参与肿瘤发生、发展进程,是恶性肿瘤的潜在治疗靶点。目的:探讨XPO1在人胰腺癌细胞中的表达、定位以及XPO1抑制剂KPT-330对肿瘤细胞增殖和凋亡的影响。方法:以蛋白质印迹法检测6株人胰腺癌细胞株和2株人永生化正常胰腺导管上皮细胞株中的XPO1表达,选择XPO1表达量最高的人胰腺癌细胞株MIA PaC a-2进行后续实验。予MIA PaC a-2细胞不同浓度KPT-330(0. 03、0. 3、3μmol/L)或DMSO处理,免疫荧光实验检测XPO1在细胞中的分布,CCK-8实验和克隆形成实验检测细胞增殖,流式细胞分析检测细胞周期和细胞凋亡,蛋白质印迹法检测XPO1和凋亡相关蛋白caspase-3、PARP表达变化。结果:XPO1主要聚集分布于MIA PaC a-2细胞的核膜,经KPT-330处理后,XPO1的核膜高聚现象消失。与DMSO组相比,KPT-330可抑制MIA PaC a-2细胞的XPO1表达,并能抑制细胞增殖,诱导细胞凋亡,作用呈浓度依赖性。机制研究显示KPT-330可诱导MIA PaC a-2细胞发生细胞周期S期阻滞,上调cleavedcaspase-3、cleaved-PARP表达。结论:XPO1抑制剂KPT-330可能通过调节细胞周期分布、诱导细胞凋亡而抑制人胰腺癌细胞增殖。
Background:XPO1(exportin1),the main mediator of nucleocytoplasmic transport,is overexpressed in a variety of human malignancies and closely related to the occurrence and development of tumor.Thus,XPO1might be served as a potential therapeutic target for the treatment of cancer.Aims:To investigate the expression and localization of XPO1in human pancreatic cancer cells and the effect of XPO1inhibitor KPT-330on proliferation and apoptosis of cancer cells.Methods:Expression of XPO1was determined in6human pancreatic cancer cell lines and2human immortalized pancreatic ductal epithelial cell lines by Western blotting.MIA PaCa-2was identified as the XPO1most up-regulated human pancreatic cancer cell line for further investigation.MIA PaCa-2cells were treated with DMSO or KPT-330at different concentrations(0.03,0.3,and3μmol/L).Cellular localization of XPO1was detected by immunofluorescent assay;cell proliferation was assessed by CCK-8assay and colony formation assay;cell cycle and apoptosis were analyzed by flow cytometry;Western blotting was performed to reveal the changes of XPO1and apoptosis-related proteins(caspase-3and PARP).Results:XPO1was mainly located on the nuclear envelope of MIA PaCa-2cells,and KPT-330treatment diminished the high aggregation of XPO1on nuclear envelope.Compared with cells treated with DMSO,KPT-330reduced the expression of XPO1,inhibited cell proliferation and induced apoptosis of MIA PaCa-2cells in a dose-dependent manner.Furthermore,S-phase arrest as well as upregulation of cleaved-caspase-3and cleaved-PARP were observed in KPT-330-treated MIA PaCa-2cells.Conclusions:XPO1inhibitor KPT-330can significantly inhibit the proliferation via regulating cell cycle and enhance the apoptosis of human pancreatic cancer cells.
作者
李苑华
庄燕妍
黄娴娴
陈文颖
黄凤婷
张世能
LI Yuanhua;ZHUANG Yanyan;HUANG Xianxian;CHEN Wenying;HUANG Fengting;ZHANG Shineng(Department of Gastroenterology,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120)
出处
《胃肠病学》
2018年第11期658-663,共6页
Chinese Journal of Gastroenterology
基金
国家自然科学基金面上项目(81572348)
广东省科技计划项目(2016A020215060)
