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| 人肝癌PLC/PRF-5细胞中干细胞样细胞的分离及其特异性miRNAs的筛选 | |
| 引用本文： | 郑秋红,许扬梅,魏植强,龚福生,杨建伟,谢云青,应敏刚. 人肝癌PLC/PRF-5细胞中干细胞样细胞的分离及其特异性miRNAs的筛选[J]. 中国肿瘤生物治疗杂志, 2011, 18(2): 126-132. DOI: 10.3872/j.issn.1007-385X.2011.02.002 |
| 作者姓名： | 郑秋红 许扬梅 魏植强 龚福生 杨建伟 谢云青 应敏刚 |
| 作者单位： | 1. 福建医科大学附属教学医院,福建省肿瘤医院肿瘤分子生物学研究室,福建,福州,350014 2. 福建医科大学附属教学医院福建省肿瘤医院肿瘤内科,福建,福州,350014 3. 福建医科大学附属教学医院福建省肿瘤医院钟瘤外科,福建,福州,350014 |
| 基金项目： | 福建省自然科学基金资助项目 (No.2008J0077) |
| 摘 要： | 目的:分选及鉴定人肝癌PLC/PRF-5细胞中的肝癌干细胞样细胞,研究其microRNAs(miRNAs)表达谱。方法:以ABCG2为表面标志,免疫磁珠法分选、流式细胞术检测ABCG2+和ABCG2-PLC/PRF-5细胞,观察ABCG2+与ABCG2-PLC/PRF-5细胞的琼脂克隆形成能力和接种NOD/SCID小鼠的成瘤能力。应用miRNA芯片筛选ABCG2+和ABCG2-PLC/PRF-5细胞差异表达的miRNAs,real-time PCR验证部分差异表达的miRNAs。结果:免疫磁珠分选的ABCG2+PLC/PRF-5细胞纯度可达(84.20±4.52)%。ABCG2+PLC/PRF-5细胞比ABCG2-PLC/PRF-5细胞形成更多、更大的克隆集落(47.17±10.50 vs23.33±7.31,P<0.05);NOD/SCID小鼠接种1×104个ABCG2+PLC/PRF-5细胞即可成瘤,而ABCG2-PLC/PRF-5细胞至少需要5×105个才可成瘤;5×105个细胞时,ABCG2+PLC/PRF-5细胞组的肿瘤体积显著大于ABCG2-PLC/PRF-5细胞组[(3.73±1.19)cm3 vs(0.72±0.57)cm3,P<0.01]。ABCG2+PLC/PRF-5细胞和ABCG2-PLC/PRF-5细胞差异表达的miRNAs有20个:上调的13个,下调的7个;real-time PCR验证其中的hsa-miR-30a和hsa-miR-630的差异表达,其结果与miRNA芯片结果基本一致。结论:人肝癌细胞系PLC/PRF-5中ABCG2+细胞具有肿瘤干细胞的特性;ABCG2+和ABCG2-PLC/PRF-5细胞差异表达的miRNAs有20个,它们在肝癌发病中可能起重要的调控作用。 |
| 关 键 词： | 肝癌 肝癌干细胞 microRNA 表达谱 免疫磁珠分选 |
| 收稿时间： | 2011-01-23 |
| 修稿时间： | 2011-03-25 |
Isolation of stem-like cells from human hepatocellular carcinoma PLC/PRF-5 cells and screening for their specific miRNAs |
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| ZHENG Qiu-hong,XU Yang-mei,WEI Zhi-qiang,GONG Fu-sheng,YANG Jian-wei,XIE Yun-qing and YING Min-gang. Isolation of stem-like cells from human hepatocellular carcinoma PLC/PRF-5 cells and screening for their specific miRNAs[J]. Chinses Journal of Cancer Biotherapy, 2011, 18(2): 126-132. DOI: 10.3872/j.issn.1007-385X.2011.02.002 | |
| Authors: | ZHENG Qiu-hong XU Yang-mei WEI Zhi-qiang GONG Fu-sheng YANG Jian-wei XIE Yun-qing YING Min-gang |
| Affiliation: | Department of Molecular Biology of Tumor, Affiliated Teaching Hospital of Fujian Medical University, Tumor Hospital of Fujian Province, Fuzhou 350014, Fujian, China |
| Abstract: | Objective: To isolate stem-like cells from hepatocellular carcinoma cell line PLC/PRF-5 and to study their miRNA profile.Methods:ABCG2+ and ABCG2-PLC/PRF-5 cells were isolated from the PLC/PRF-5 cell line by magnetic activated cell sorting(MACS) method,and further identified by flow cytometry.The colony formation ability in soft agar and tumor formation ability in NOD/SCID mice of ABCG2+ and ABCG2-PLC/PRF-5 cells were observed.miRNA chip was adopted to screen the differentially expressed miRNAs between ABCG... |
| Keywords: | hepatocellular carcinoma hepatocellular carcinoma stem cell microRNA profile magnetic activated cell sorting |
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