基于SP细胞分选法初步鉴定卵巢癌干细胞表面标志_临床医学论文.docVIP

基于SP细胞分选法初步鉴定卵巢癌干细胞表面标志_临床医学论文 基于SP细胞分选法初步鉴定卵巢癌干细胞表面标志_临床医学论文 作者：张弦　蒋翠莲　王宝偲　刘蔷　左鹏飞　赵枫姝　窦骏 【摘要】 目的： 基于侧群（SP）细胞分选方法，初步鉴定卵巢癌干细胞（CSC) 表面标志，为临床靶向治疗卵巢癌提供靶分子。方法： 检测、分离卵巢癌细胞株A2780中的SP细胞，并对SP与Non细胞作细胞生物学鉴定，包括克隆形成能力、致瘤能力、耐药性、定量ATP结合框蛋白家族G2蛋白 (ABCG2)检测等；进一步用免疫磁选方法筛出ABCG2+及ABCG2-细胞，同样作上述细胞生物学鉴定，探讨ABCG2分子能否作为卵巢CSC的表面标志。结果： 卵巢癌A2780细胞株中的SP细胞在体外强克隆形成能力、裸鼠体内的高致瘤性、对化疗药物长春新碱较强耐受性及高表达ABCG2分子等生物学特性基本符合CSC的特征。ABCG2＋细胞的克隆增殖能力略强于ABCG2-细胞；ABCG2+细胞的耐药性与SP细胞的耐药性一致。结论： A2780细胞株中的SP细胞基本符合CSC的生物学特征，其高表达的ABCG2分子参与维持SP细胞表型，特别是多药耐药性。ABCG2分子可作为靶向治疗CSC的靶分子，逆转肿瘤的多药耐药性。 【关键词】 侧群细胞　卵巢癌　癌干细胞　ABCG2分子　靶向治疗　 Abstract： Objective To explore a target molecule for the clinical targeted therapy of ovarian cancer. Methods The SP cells and non SP cells were isolated respectively from the human ovarian cell 2780 strain by fluorescence activated cell sorting (FACS) and their biological characteristics were identified by analysis of their ability to form colonies in common media, tumorigenicity in Balb/c nude mice, and of their resistance to chemotherapeutic agents vinblastin. The expression of ABCG2 was detected with realtime quantitative RTPCR (qRTPCR). The specific ABCG2 phenotype cells were further isolated from the ovarian cell A2780 strain for the same identification as above by the monoantibody labeled with immune magnetic beads by magnetic activated cell sorting system. Results The SP cells proliferative potency, clone formation ability in the common media, the tumorigenicity in Balb/c nude mice, and resistance to vinblastin in vitro were basically coincident with characteristics of cancer stem cells（CSC）. The ABCG2＋cells proliferative potency and resistance to vinblastin in vitro were higher than those of the ABCG2-cells. Conclusion The SP cells in the ovarian cell 2780 strain possess the traits of ovarial CSC and expressed ABCG2 molecule maintains the SP cells phenotype, especially in multidrug resistance. The ABCG2 molecule may be responsible for target molecule of the targeted therapy of ovarian canc