中国肿瘤生物治疗杂志2011，Vol.18Issue(2)：126-132,7.DOI:10.3872/j.issn.1007-385X.2011.02.002
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
Isolation of stem-like cells from human hepatocellular carcinoma PLC/PRF-5 cells and screening for their specific miRNAs
摘要
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 ABCG2 + and ABCG2 - PLC/PRF-5 cells; and real-time PCR assay was used to confirm the results of miRNA chip. Results:The purity of ABCG2 + PLC/PRF-5 cells isolated by MACS method was ( 84.20 ± 4.52 ) ％. The colony number and size formed by ABCG2 + PLC/PRF-5 cells were more and larger than those formed by ABCG2 - cells (47.17 ± 10.50 vs 23.33 ±7.31, P ＜0.05). 1 × 104 ABCG2 + cells could form tumors with at least 5 × 105 cells needed for ABCG2 - cells. The size of tumors generated by 5 × 105 ABCG2 + cells was larger than that by the ABCG2 - cells ( [ 3.73 ± 1.19 ]cm3 vs [0.72 ± 0.57 ] cm3, P ＜ O. Ol ). There are 20 miRNAs differentially expressed between ABCG2 + and ABCG2- cells, with 13 up-regulated and 7 down-regulated. Realtime PCR assay was applied to further verify the differential expression of hsa-miR-30a and hsa-miR-630, and the results were in accordance with those of miRNA chip. Conclusion: ABCG2+ PLC/PRF-5 cells in hepatocellular carcinoma cell line PLC/PRF-5 have the properties of cancer stem cells. Twenty miRNAs are differentially expressed between ABCG2 + and ABCG2 - PLC/PRF-5 cells, which might play important roles in the carcinogenesis of hepatocellular carcinoma.关键词
肝癌/肝癌干细胞/microRNA/表达谱/免疫磁珠分选分类
生物科学引用本文复制引用
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):126-132,7.基金项目
福建省自然科学基金资助项目(No.200810077). （No.200810077）
