肠癌细胞系SW-620中肿瘤干细胞相关SP亚群分析  

Analysis of tumor stem cells-related side population in intestine cancer cell line SW-620

作　　者：高钢龙[1] 张学利[1] 刘文勇[2] 张萍[3] 叶冬霞[3] 

机构地区：[1]南方医科大学附属奉贤医院普外科(上海市奉贤区中心医院普外科),上海市201400 [2]上海交通大学医学院附属第九人民医院普外科,上海市200011 [3]上海交通大学医学院附属第九人民医院口腔肿瘤实验室,上海市200011

出　　处：《中国组织工程研究与临床康复》2010年第36期6790-6793,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：上海市卫生局科学基金资助项目(2007157);课题名称"分离鉴定人胃肠道癌中肿瘤干细胞相关的SP亚群的初步研究"~~

摘　　要：背景:在某些恶性肿瘤中,SP细胞具有肿瘤干细胞的特性,如人脑瘤、乳腺癌等肿瘤干细胞的研究就是采用了这种方法。目前关于肠癌干细胞的研究尚处于探索阶段,国内外尚无肠癌干细胞分离、鉴定成功的报道。目的:分析肠癌细胞系SW-620中是否包含肿瘤干细胞相关的SP亚群。方法:制备SW-620细胞悬液,经Hoechst33342和PI染色,实验组加入Hoechst33342至终浓度为5mg/L,维拉帕米组同时加入维拉帕米至终浓度5mg/L,流式细胞仪分析SP亚群,共选取3次实验的结果。结果与结论:通过Hoechest33342蓝光和红光双参图,实验组SP细胞亚群位于左下角两种荧光均阴性或很弱的区域,经3次检测,肠癌细胞株SW620中SP比例分别为0.1%,1.0%和0.5%,经维拉帕米阻断后SP细胞比例均为0。提示人肠癌细胞系SW620中存在SP亚群细胞,即提示肠癌干细胞存在的可能;维拉帕米可抑制染料外从排而明显减少SP细胞的比例。BACKGROUND：In some malignant tumors, side population （SP） cells have the property of tumor stem cells.The studies of human cerebroma and breast carcinoma used this method.At present, studies concerning intestine cancer stem cells are in exploration stage.There is no report on successful isolation and identification of intestine cancer stem cells.OBJECTIVE：To identify the SP of cancer stem-like cells in intestine cancer cell line SW-620.METHODS：SW-620 tumor cells suspension was prepared and stained with Hoechst33342 and propidium iodide.In the experimental group, Hoechst33342 was added until the final concentration was 5 mg/L.In the verapamil group, an additional verapamil was added until the final concentration was 5 mg/L.SP was analyzed using flow cytometry.Results from three tests were selected.RESULTS AND CONCLUSION：Using Hoechst33342 blue and red light double reference graph.In the experimental group, SP cell line located at the lower left quarter region where two kinds of fluorescence were negative or very weak.Following three tests, SP proportion in SW620 was respectively 0.1%, 1.0% and 0.5%.Following verapamil treatment, the SP cell proportion was 0.These indicated that SW620 cell line contains SP cells; i.e.the possibility of SW620 cell line existence.Verapamil can inhibit dye, and obviously decrease the proportion of SP cells.

关 键 词：肿瘤干细胞 SW-620细胞 SP亚群 肠癌 细胞鉴定 

分 类 号：R394.2[医药卫生—医学遗传学]