卵巢癌细胞系ID8中肿瘤干细胞的分离及生物学特性鉴定

Isolation and biological identification of tumor stem cells from ovarian cancer ID8 cell lines

背景:肿瘤干细胞在肿瘤发生和发展中起着至关重要的作用,但鼠卵巢癌细胞系ID8中是否存在肿瘤干细胞仍未见报道。目的:分选小鼠源性卵巢癌细胞系ID8的肿瘤干样细胞,鉴定其生物学特性。方法:应用无血清培养基培养ID8细胞,将获得悬浮球细胞传代扩增。体外采用CCK8法、平板克隆、Transwell小室侵袭及体外药敏实验观察悬浮球细胞与常规培养的ID8细胞在生物学功能上的差异。应用定量PCR以及Western blot检测悬浮球细胞和非悬浮球细胞的肿瘤干细胞标志ALDH1的表达水平,并通过小鼠致瘤实验观察其成瘤能力。结果与结论:小鼠卵巢癌ID8细胞能在无血清培养基中形成可稳定传代的悬浮球细胞,与非悬浮球细胞相比,具有较强的增殖、自我更新、侵袭、耐药及致瘤的能力,并高表达肿瘤干细胞标志ALDH1。说明无血清培养法可以有效的富集具有卵巢癌干样细胞特征的ID8悬浮球细胞。

BACKGROUND:Tumor stem cells play an important role in the initiation and development of tumor.However there is no report on whether tumor stem cells exist in mouse ovarian cancer cell line ID8.OBJECTIVE:To sort tumor stem-like cells from murine ID8 ovarian cancer cell line and to investigate its biological characteristics.METHODS:ID8 cells were cultured in the serum-free medium to derive tumor spheres.Cell counting kit 8,plate colony formation,Transwell assays and chemosensitivity testing were adopted to analyze the biological differences between suspending sphere cells and conventional cultured ID8 cells.Real-time PCR and western blot assays were performed to examine the expression of ALDH1.A mouse model of tumorigenicity was used to observe the tumorigenesis in vivo.RESULTS AND CONCLUSION:Ovarian ID8 cells grew into stable suspending tumor spheres in the serum-free medium.Compared with conventional ID8 cells,suspending sphere cells had stronger abilities of proliferation,self-renewal,invasion,drug resistance and tumorigenesis.Moreover,ALDH1 expression in suspending spheres cells was higher than that in conventional cultured ID8 cells.These findings indicate that serum-free medium culture can effectively enrich ID8 suspending sphere cells which have the properties of ovarian cancer stem cells.