耐吉西他滨胰腺癌细胞株SW1990/GZ上皮间质转化现象的研究  

Investigation of epithelial-mesenchymal transition in gemcitabine-resistant cancer cell line SW1990/GZ

作　　者：王辉[1] 张烨[1] 吴峻立[1] 徐泽宽[1] 苗毅[1] 

机构地区：[1]南京医科大学第一附属医院普外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2012年第3期306-309,373,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(30972912)

摘　　要：目的:观察耐吉西他滨胰腺癌细胞株(SW1990/GZ)上皮间质转化的现象。方法:倒置显微镜下观察亲代SW1990细胞与SW1990/GZ细胞的形态差别;流式细胞仪检测细胞周期;免疫印迹法(Western blot)检测上皮间质转化相关蛋白表达;Transwell小室测定细胞侵袭迁移能力;通过表面特异抗原(CD44+、CD24+和CD133+)标记流式细胞仪检测肿瘤干细胞比例。结果:在形态学上,SW1990/GZ表现出明显的间质细胞特征,与亲代SW1990相比处于G1期的细胞无明显差异;SW1990/GZ细胞低表达上皮细胞标志物E-cadherin,而高表达间质细胞的标记物Vimentin;SW1990/GZ细胞侵袭和迁移能力明显增强(P<0.01);与亲代细胞SW1990相比,SW1990/GZ细胞中CD44+CD24+细胞比例以及CD133+细胞比例分别提高了2倍和4倍以上(P<0.01)。结论:SW1990耐吉西他滨细胞株(SW1990/GZ)发生了明显的上皮间质转化过程,肿瘤干细胞比例明显增多。Objective：To explore epithelial-mesenchymal transition（EMT） in gemcitabine-resistant cancer cell line SW1990/GZ.Methods： Morphological characteristics of SW1990 cells and SW1990/GZ cells were observed by inverted microscope.Cell cycle distribution was analyzed by flow cytometry.The expressions of EMT-associated markers were determined by Western blot.Invasion and migration of cells were detected by transwell.The proportion of cancer stem cells was analyzed by flow cytometry after labeled the surface specific antigen.Results： SW1990/GZ cells exhibited mesenchymal morphology,and no significant G1 accumulation phenotype in SW1990/GZ cells compared with SW1990 cells.SW1990/GZ cells reduced expression of the epithelial marker E-cadherin but increased expression of the mesenchymal marker vimentin.SW1990/GZ cells have greater invasion and migration activity（P 0.01）.Compared with SW1990 cells,the number of CD44＋CD24＋ cells and CD133＋ cells increased by 2 times and 4 times in SW1990/GZ cells（P 0.01）.Conclusion：SW1990/GZ cells have undergone an EMT,and are enriched with cancer stem cells.

关 键 词：胰腺癌 上皮间质转化 肿瘤干细胞 侵袭 迁移 

分 类 号：R735.9[医药卫生—肿瘤]