乙醛脱氢酶1作为喉癌Hep-2细胞系肿瘤干细胞标志物的实验研究

目的 探讨乙醛脱氢酶1(ALDH1)在人喉癌Hep-2细胞系中的表达,观察ALDH1高表达细胞的体外生长特性,确定喉癌Hep-2细胞系肿瘤干细胞标志物.方法 采用荧光细胞化学染色和流式细胞术观察和检测喉癌Hep-2细胞系中的ALDH1的表达.采用流式分选术分离ALDH1高表达细胞,以四甲基偶氮唑蓝(MTT)法观察ALDH1不同亚群细胞的体外增殖能力,以流式细胞术检测ALDH1高表达细胞的体外分化能力.采用裸鼠成瘤实验比较不同亚群细胞的成瘤能力.结果 喉癌Hep-2细胞系中ALDH1呈不同程度的表达,其中ALDH1高表达细胞占2.9％±0.6％.ALDH1高表达细胞的体外增殖能力高于ALDH1低表达和未分选细胞.在含血清的培养液中,经过6d培养,ALDH1高表达细胞由最初分选后的94.2％±3.8％下降至分选前水平.ALDH1高表达细胞的成瘤能力显著高于其他亚群.结论 喉癌Hep-2细胞系中,ALDH1高表达细胞具有很强的体外分化能力、增殖能力和成瘤能力,可以作为Hep-2细胞系肿瘤干细胞的标志物之一.

Aldehyde dehydrogenase 1 can be used as a new marker of cancer stem cells in laryngeal cancer cells in vitro

Objective To detect the expression of aldehyde dehydrogenase 1 in human laryngeal cancer cells in vitro,and to explore whether it can be used as a marker of stem cells in human laryngeal cancer.Methods Fluorescence staining and flow cytometry were used to detect the expression of ALDH1 in a human laryngeal cancer Hep-2 cell line,and fluorescence activated cells sorting was used to separate ALDH1br cells.ALDH1 tumor cells were cultured and their ability of proliferation and differentiation was observed in vitro.Results The expression of ALDH1 in Hep-2 cells was different.The number of cells highly expressing ALDH1 was 2.9％ ± 0.6％.Compared with ALDH1 low cells and unsorted cells,ALDH1 br cells exibited increased proliferation ability.In serum-containing RPM I1640 culture medium,the proportion of ALDH1br cells was decreasing as days passed.The percentage of ALDH1br cells decreased from 94.2％ ±3.8％ to the level before sorting. The ALDH1br cells demonstrated enhanced tumorigenic ability in nude mice.Conclusions In the laryngeal cancer Hep-2 cell line,the highly ALDH1-expressing cells show a strong ability of differentiation,proliferation and tumorigenesis.It indicates that ALDH1 can be used as a new marker of stem cells of laryngeal cancer cells.