基于ABCG2的卵巢癌干样细胞分选及体外生物学研究  

Using ABCG2-molecule-expressing side population cells to identify cancer stem-like cells in a human ovarian cell lines

作　　者：龚丽华[1] 朱含笑 周海虹 石智[2] 颜笑健[1] GONG Lihua;ZHU Hanxiao;ZHOU Haihong(Department of Gynecology,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第一医院妇科,325000 [2]暨南大学细胞生物学与生物医学研究所、生命科学与技术学院

出　　处：《浙江医学》2020年第11期1106-1111,I0003,共7页Zhejiang Medical Journal

基　　金：国家自然科学基金青年科学基金项目(81503293)。

摘　　要：目的探讨卵巢癌干样细胞(OCSCs)的生物学特点,分析OCSCs与卵巢癌细胞三磷酸腺苷结合盒跨膜转运蛋白G超家族成员2(ABCG2)表达的相关性,阐述两者与卵巢癌化疗抗药的关系。方法采用ABCG2特异性抑制剂FTC作为侧群法流式细胞分选对照组的抑制剂,分选并检测卵巢癌细胞株中侧群(SP)细胞比例。体外克隆形成实验和体外分化实验研究细胞增殖、分化等生物学特点。噻唑蓝法检测卵巢癌SP细胞与非侧群(NSP)细胞对常用卵巢癌化疗药物的敏感性,并检测卵巢癌细胞株ABCG2+细胞比例。结果卵巢癌细胞中存在SP细胞,分选纯度达95%左右,其中SP细胞比例在0.19%~4.88%。SP细胞具有体外分化能力,且体外克隆形成能力明显高于NSP细胞。卵巢癌SP细胞显示抗药性,顺铂、紫杉醇、吉西他滨及多柔比星对SP细胞的半数抑制浓度明显高于NSP细胞(均P<0.05)。卵巢癌细胞中SP细胞比例与ABCG2+细胞比例呈正相关。结论OCSCs具有较强的增殖和体外分化、克隆形成能力,对化疗药物抗药性强。OCSCs比例与耐药蛋白ABCG2存在正相关性,或为卵巢癌治疗中逆转耐药提供新的靶点。Objective To identify ovarian cancer stem cells(OCSCs)in human ovarian cell lines and to investigate the biological characteristics of OCSCs.Methods The side population(SP)and non-side population(NSP)cells were sorted from human ovarian cancer cell lines by fluorescence-activated cell sorting(FACS).The proportion of SP cells in ovarian cancer cell line was determined by using FTC as an inhibitor for the control group.In vitro clone formation and differentiation experiment were used to study the biological characteristics of cell proliferation and diffe-rentiation.MTT assay was used to detect the sensitivity of SP cells and NSP cells to common chemotherapy drugs for ovarian cancer,and the proportion of ATP-binding cassette subfamily G member 2(ABCG2+)cells in ovarian cancer cell line was also detected.Results There were SP cells in ovarian cancer cells.The purity of SP cells was 95%,and the proportion of SP cells was 0.19%~4.88%.SP cells had the ability of differentiation in vitro,and the ability of clone formation in vitro was significantly higher than that of NSP cells.In addition,the IC50 of cis-platinum,Taxol,gecitabine and doucirocin in SP were all much higher than in NSP(P<0.05).The proportion of SP cells in ovarian cancer cells was positively correlated with the proportion of ABCG2+cells.Conclusion Ovarian cancer stem cells have strong ability to proliferate,differentiate and clone in vitro.The proportion of cancer stem-like cells in ovarian cancer is positively correlated with drug-resistant protein ABCG2,which may provide a new target for the reversal of drug resistance in the treatment of ovarian cancer.

关 键 词：卵巢癌干样细胞 侧群细胞 三磷酸腺苷结合盒跨膜转运蛋白G超家族成员2 化疗耐药 

分 类 号：R737.31[医药卫生—肿瘤]