宫颈癌细胞系中干细胞球样细胞的筛选与鉴定研究

Isolation and identification of sphere-forming stem-like cells from the cervical cancer cell line

目的探讨从人宫颈癌细胞系Ca Ski中分离培养宫颈癌干细胞球样细胞的可行性,并观察其体外生物学特性。方法采用肿瘤干细胞无血清悬浮培养法富集Ca Ski细胞系球样细胞,通过体外传代培养、添加血清诱导分化等技术鉴定其干性,利用流式细胞技术检测细胞周期,并应用蛋白印记法及细胞免疫荧光等方法从分子水平检测相关因子的表达。结果从Ca Ski细胞系中分离到的干细胞球样细胞可以在无血清培养基中悬浮生长成细胞球,并可在体外稳定传代,具有自我更新能力和分化潜能;肿瘤干细胞球样细胞多处于G0/G1期。与其他分子标志物相比,干细胞球样细胞的CD44明显高表达;与亲本细胞相比,干细胞球样细胞的干性相关因子SOX2、OCT4、β-catenin表达较高。结论利用肿瘤干细胞无血清悬浮培养法可以从Ca Ski细胞系中分离富集到具有稳定肿瘤干细胞性质的细胞群。

Objective To isolate and identify sphere-forming stem-like cells from cervical cancer cell line（Ca Ski）, and observe its biological characteristics in vitro. Method We enriched sphere- forming stem- like cells using sphere suspension culture system, and confirmed its stem cell- associated characteristics by evaluating the renewal and serum-induced differentiation abilities. Western Blot technology was applied to detect the expression of stem cell-related factors at the protein level. Result The sphere- forming stem- like cells isolated from Ca Ski can grow into spheres in suspension in serum-free medium, and were able to self-renew. Ca Ski cancer stem cell-like cells were enriched in the G0/G1 phase. Compared with other stem cell markers, CD44 has a high expression and can be as the useful marker for cervical cancer cells. Compared with its parental cells, SOX2, OCT4, β- catenin has a high expression of stem cell- related factors. Conclusion We successfully isolated and enriched sphere- forming stem- like cells from cervical cancer cell line Ca Ski, using serum-free sphere suspension culture system.