基础研究(Basic Research)

胆管癌干细胞的分离、培养及鉴定

Published at: 2016年第25卷第2期

王晓博 1 , 邓小峰 1 , 朱建伟 1 , 刘志鹏 1 , 马望 2 , 耿艺维 2 , 熊力 1 , 文宇 1 , 苗雄鹰 1
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
通讯作者 雄鹰 苗 Email: 362802445@qq.com
DOI: 10.3978/j.issn.1005-6947.10.3978/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。


Isolation, cultivation and identification of cholangiocarcinoma stem cells

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.


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引用

引用本文: 晓博 王, 小峰 邓, 建伟 朱, 志鹏 刘, 望 马, 艺维 耿, 力 熊, 宇 文, 雄鹰 苗. 胆管癌干细胞的分离、培养及鉴定[J]. 中国普通外科杂志, 2016, 25(2): 219-224.
Cite this article as: WANG Xiaobo, DENG Xiaofeng, ZHU Jianwei, LIU Zhipeng, MA Wang, GENG Yiwei, XIONG Li, WEN Yu, MIAO Xiongying . Isolation, cultivation and identification of cholangiocarcinoma stem cells [J]. Chin J Gen Surg, 2016, 25(2): 219-224.