神经生长因子受体阳性的人食管鳞癌细胞的分选及其干细胞特性

 目的 应用免疫磁珠分选(magnetic activated cell sorting,MACS)人食管鳞癌细胞(esophageal squamous carcinoma cell,ESCC)中的神经生长因子受体(nerve growth factor receptor,NGFR)阳性细胞,并对其肿瘤干细胞样特征进行研究。方法 采用免疫磁珠分选技术分选人食管鳞癌细胞系中的NGFR阳性细胞,然后通过流式细胞仪法测定分选细胞的纯度及细胞周期分布,并计算细胞回收率;MTT法测定细胞数,绘制生长曲线并计算细胞群体倍增时间;平板克隆实验测定平板克隆形成率;裸鼠接种成瘤实验测定体内致瘤性,从而鉴定NGFR阳性ESCC的干细胞特性。结果 NGFR阳性细胞约占肿瘤细胞的1.89%,经过磁珠分选后,其纯度显著提高,达89.94%,细胞回收率为19.83%。与未分选细胞相比,NGFR阳性细胞的增殖速度明显提高,生长曲线左移,细胞倍增时间缩短［(25.22±8.15)h vs. (47.41±12.69)h,P<0.05］,平板克隆形成率显著增加(16.27%±8.31% vs. 1.22%±0.35%,P<0.05),处于活跃增殖期S、G2、M期的细胞显著增加(72.7% vs. 18.8%,P=?);裸鼠皮下接种的转移瘤形成率增加(8/10 vs. 1/10,P=?),转移瘤重量显著增加［0.72±0.53)g vs. (0.16±0.26)g,P<0.05］。结论 应用免疫磁珠分选技术能有效分选出人食管鳞癌细胞中的NGFR阳性细胞,NGFR阳性细胞具有肿瘤干细胞特性。

Isolation of nerve growth factor receptor positive cells from human esophageal squamous carcinoma cells and their biological properties

Objective To isolate nerve growth factor receptor(NGFR) positive cells from human esophageal squamous carcinoma cells(ESCC) and to observe their biological properties.Methods NGFR positive cells were isolated from human ESCCs by magnetic activated cell sorting(MACS).NGFR positive rate of isolated cells and cell generation cycles were detected by flow cytometry,meanwhile the cell recovery rate was calculated.Cell numbers were counted by MTT method,cell growth curves were obtained,and doubling generation time...