人上皮性卵巢癌细胞系侧群细胞的肿瘤干细胞样细胞生物学特性及膜蛋白差异表达的鉴定

目的 鉴定上皮性卵巢癌细胞侧群细胞(SP细胞)肿瘤干细胞样细胞生物学特性,并检测侧群细胞内差异蛋白质的表达.方法 流式分选上皮性卵巢癌细胞系SKOV3和A2780具有外泌Hochest33342染料生物学特性的侧群细胞,鉴定其肿瘤干细胞样细胞相关生物学特性.运用细胞培养稳定同位素标记(SILAC)的定量蛋白质组学技术,...

Identification of bio-functions of cancer stem-like cells and differentially expressed membrane proteins in side population cells from human epithelial ovarian cancer cell lines

Objective To validate the stemness bio-function of side population(SP)cells from epithelial ovarian cancer cell lines and to detect the differentially expressed cellular membrane proteins in SP cells.Methods SP cells of human ovarian cancer cell lines SKOV3 and A2780 which secreted bisbenzimide Hoechst33342 and located as side population were sorted by flow cytometry.Bio-functions of stemness of SP cells were validated.Through stable isotope labeling by amino acids in cell culture(SILAC),differentially expressed membrane proteins in SPcells were detected.Results SP cells of SKOV3 and A2780 were sorted successfully.The ratio of SP cells was about 0.5%and increased significantly through repeated sorting.SP cells differentiated into non-SP(NSP)cells.Stemness of SP cells was validated in vitro,including cell clone forming,resistance to cisplatin and paclitaxel in NOD/SCID mice.SILAC technique was used to detect differentially expressed membrane proteins in SP cells.Results showed that 1561 and 1989 differentially expressed proteins were detected respectively in SKOV3 and A2780 SP cells.Among those significantly up-regulated proteins in SKOV3 SP cells,12 proteins were also up-regulated in A2780 SP cells.On the contrary,13 proteins were down-regulated both in SKOV3 and A2780 SP cells.Conclusions SP cells of human epithelial ovarian cancer cells show stemness.And differently expressed membrane proteins in SP cells were detected which would be the base of further screening surface biomarker of ovarian cancer stem-like cells.