卵巢癌干细胞的分离 、鉴定及初步生物学功能研究

摘要

目的 从人卵巢上皮细胞癌建系的SKOV3细胞中分离出CD133CD117表型卵巢癌干细胞 ,探讨其相关生物学特性.方法 利用免疫磁珠技术分选出CD133CD117癌细胞 ,继而进行平皿、软琼脂克隆形成试验 ,无血清培养试验和致瘤试验探讨其初步功能 ;采用反转录聚合酶链反应检测癌细胞Wnt-2基因表达水平.结果 表型CD133CD117癌细胞在平皿、软琼脂和无血清培养基中克隆形成能力明显强于CD133-CD117-癌细胞 ;同时表现出较强致瘤性 ;Wnt-2基因在不同表型癌细胞中存在差异表达.结论 卵巢癌细胞中存在干细胞样癌细胞 ,利用磁珠分选得到的CD133CD117癌细胞具有干细胞的相关特性 ,为卵巢癌干细胞生物学特性的研究及靶向治疗提供依据.%Objective To isolate the CD133CD117 phenotype ovarian cancer stem cells from the SKOV3 cell line of human ovarian carcinoma ,and to investigate its related biological function .Methods The CD133CD117 cancer cells were sorted through MACS technology ,then the plates ,soft agar colony formation assays ,serum-free culture and tumorigenicity test were carried out to investigate its preliminary function;RT-PCR was used to detect cancer cell Wnt-2 gene expression .Results The cloning formation capability of CD133CD117 cancer cells in petri dishes ,soft agar and serum-free medium was stronger than that in CD133-CD117- cancer cells ;meanwhile showing stronger tu-morigenicity ;the expression levels of Wnt-2 gene were different among different phenotypes of cancer cells .Conclu-sion Stem cell-like cancer cells exist in ovarian cancer cells ,sorted CD133CD117 cancer cells from ovarian cancer cells by MACS have the properties of stem cells ,which provides the scientific basis for the study of biological charac-teristics and targeted therapy of ovarian cancer stem cells .