文章摘要

胆管癌干细胞的分离、培养及鉴定

作者: 1王晓博, 1邓小峰, 1朱建伟, 1刘志鹏, 2马望, 2耿艺维, 1熊力, 1文宇, 1苗雄鹰
1 中南大学湘雅二医院 普通外科，湖南 长沙 410011
2 郑州大学第一附属医院 肿瘤科，河南 郑州 450000
通讯: 苗雄鹰 Email: 362802445@qq.com
DOI: 10.3978/.10.3978/j.issn.1005-6947.2016.02.011
基金: 国家自然科学基金资助项目, 81372628

摘要

目的：从人胆管癌细胞系QBC-939中分离具有干细胞特征的肿瘤细胞，为后续胆管癌干细胞的研究提供材料。方法：用无血清培养法及流式细胞分选法从QBC-939细胞中分离得到具有干细胞特征的CD133+EpCAMhigh干细胞样细胞，继续在无血清培养基中培养，观察其成球能力，并比较CD133+EpCAMhigh干细胞样细胞与QBC-939细胞单克隆形成率、耐药性，增殖能力，干细胞相关核转录因子OCT-4、Bmi-1、E-cadherin蛋白表达情况，以及BALB/c小鼠皮下移植后的成瘤能力。结果：在无血清培养基中，CD133+EpCAMhigh干细胞样细胞具有较强的成球能力。与QBC-939细胞比较，CD133+EpCAMhigh干细胞样细胞克隆形成率、洛铂耐药性、增殖明显增加；干细胞相关核转录因子OCT-4、Bmi-1表达明显增加，而E-cadherin表达明显降低；皮下移植瘤形成率明显增加。以上差异均有统计学意义（均P<0.05）。结论：从人胆管癌QBC-939细胞中分离的干细胞样细胞具有肿瘤干细胞特性，可用于胆管癌干细胞的研究。
关键词： 胆管肿瘤 肿瘤干细胞 肿瘤干细胞测定

Isolation, cultivation and identification of cholangiocarcinoma stem cells

Authors: 1WANG Xiaobo, 1DENG Xiaofeng, 1ZHU Jianwei, 1LIU Zhipeng, 2MA Wang, 2GENG Yiwei, 1XIONG Li, 1WEN Yu, 1MIAO Xiongying
1 Department of General Surgery, the Second Xiangya Hospital, Central South University, Changsha 410011, China
2 Department of Onclongy, the First Affiliatd Hospital, Zhengzhou University, Zhengzhou 450000, China

CorrespondingAuthor:MIAO Xiongying Email: 362802445@qq.com

Abstract

Objective: To isolate from human cholangiocarcinoma QBC-939 cells, the tumor cells with stem cell properties and provide material for subsequent investigations on cholangiocarcinoma stem cells. Methods: By means of serum-free culture and fluorescence-activated cell sorting, the CD133+EpCAMhigh stem cell-like cells with stem cell properties were isolated from QBC-939 cells, and were continuously cultured in the serum-free medium for observing their sphere forming ability. Then, the colony formation rate, drug resistance and cell proliferation ability, and the expressions of stem cell-associated nuclear transcription factors that included OCT-4, Bmi-1 and E-cadherin, as well as the tumor formation ability after subcutaneous implantation in BALB/c mice were compared between CD133+EpCAMhigh stem cell-like cells and QBC-939 cells. Results: CD133+EpCAMhigh stem cell-like cells showed a relatively strong sphere forming ability in the serum-free medium. In CD133+EpCAMhigh stem cell-like cells compared with QBC-939 cells, the colony formation rate, resistance to lobaplatin, and proliferative ability were significantly increased; the OCT-4 and Bmi-1 protein expressions were up-regulated, while the E-cadherin protein expression was down-regulated; the rate of subcutaneous xenograft formation was significantly increased. All the differences had statistical significance (all P<0.05). Conclusion: The stem cell-like cells derived from cholangiocarcinoma QBC-939 cells possess stem cell properties, and can be used for cholangiocarcinoma stem cell research.
Keywords：