肝癌肿瘤干细胞的分离鉴定及差异性小分子RNA筛选  

Isolation and Identification of Liver Cancer Stem Cells and Analysis of Differentially Expressed MicroRNAs

作　　者：许英晨[1] 管利东[1,2] 周军年[1] 曾泉[1] 袁红丰[1] 李思霆[1] 管兆轩[1] 何丽娟[1] 南雪[1] 陈琳[1] 岳文[1] 裴雪涛[1] 

机构地区：[1]军事医学科学院野战输血研究所干细胞与再生医学研究室,北京100850 [2]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国生物工程杂志》2013年第1期1-7,共7页China Biotechnology

基　　金：国家"863"计划(2011AA020109);国家重点基础研究发展规划(重大科学研究计划)(2011CB964804);国家自然科学基金(30971145;81101812)资助项目

摘　　要：旨在分离并鉴定肝癌肿瘤干细胞,并对其异常表达的microRNA进行了初步筛选。首先利用流式细胞术及免疫荧光技术分别在肝癌细胞系及肝癌组织标本中确认了CD90+细胞的存在;随后利用免疫磁珠分选技术从肝癌细胞系MHCC97-H中分离出了CD90+细胞,化疗药物耐药性实验表明,CD90+细胞具有明显的化疗药物耐受能力;两次裸鼠皮下成瘤性实验也表明CD90+细胞具有较强的成瘤能力,而CD90-细胞不具备成瘤能力。上述实验充分说明:CD90+细胞具有肝癌肿瘤干细胞特性。因此利用该细胞模型进行了肝癌肿瘤干细胞CD90+细胞和非肝癌肿瘤干细胞CD90-细胞之间差异表达的microRNA的检测,结果表明,CD90+细胞与CD90-细胞之间共有92个microRNAs的表达存在差异性,其中在CD90+细胞中高表达的microRNAs有52个;在CD90-细胞中高表达的microRNAs有40个。The aim of the study is to isolate and identify liver cancer stem cells and screen the differentially expressed microRNAs.CD90＋ cells were confirmed the existence in eight hepatocellular carcinoma cell lines and specimens by flow cytometry and immunofluorescence.Next,CD90＋ cells isolated from MHCC97-H cell line by MACS.Drug chemoresistance assay showed CD90＋ cells have a stronger drug resistance capacity than CD90-cells.CD90＋ cells also have a stronger ability to initiate tumor formation than CD90-cells in a xenograft model in nude mice.Taken together,liver cancer stem cells may be enriched in CD90＋ populations.Therefore,we examined the differentially expressed microRNAs between CD90＋ cell and CD90-cells by means of microRNA chip.It is revealed that 92 microRNAs are differentially expressed between CD90＋ cell and CD90-cells,of which 52 microRNAs are highly expressed in CD90＋ cells and 40 microRNAs are highly expressed in CD90-cells.

关 键 词：肝癌 肿瘤干细胞 CD90 小分子RNA 

分 类 号：R394.2[医药卫生—医学遗传学]