目的 利用干细胞中乙醛脱氢酶（ALDH）高表达的特性分选并鉴定肿瘤细胞中存在的ALDH high 、ALDH low 及ALDH neg 细胞亚群。 方法 以流式细胞术检测小鼠肝癌H22、人胃癌BGC-823、结肠癌LS-174T、肝癌SMMC-7721细胞及胰腺癌SW-1990和Panc-1细胞系中ALDH+细胞的表达；在确定ALDH阳性表达的细胞中，以ALDH表达的高低差异，将细胞分选并定义为ALDH high 、ALDH low 、ALDH neg 细胞亚群；对分选出的3种细胞亚群进行CD133/CD44表型及细胞周期分析。 结果 （1）在小鼠肝癌H22细胞、人结肠癌LS-174T细胞、胰腺癌SW-1990细胞和Panc-1细胞中存在着ALDH阳性表达，而在人胃癌BGC-823细胞和肝癌SMMC-7721细胞中未见ALDH表达；（2）在小鼠肝癌H22细胞中能明确分选出存在荧光差异的3群细胞即ALDH high 、ALDH low 和ALDH neg 细胞亚群；（3）H22的ALDH high 、ALDH low 、ALDH neg 3个亚群中CD133/CD44表达存在差异，其表达比例依次减少；（4）H22细胞中ALDH high 亚群主要处于G 0/G 1期，ALDH low 亚群处于S期，而ALDH neg 亚群处于G 2/M期。 结论 在肿瘤细胞中存在着介乎于肿瘤干细胞与终末肿瘤细胞之间的特异性亚群，该亚群为肿瘤治疗提供了新的方向。
Abstract
Objective To define the contents of different stem characteristic subsets in tumor cells based on the high expression of ALDH. Methods We identified the ALDH expression by FCM in cell line cells of Kunming mouse liver cancer H22, gastric cancer BGC-823, colon cancer LS-174T, liver cancer SMMC-7721, pancreatic cancer SW-1990 and Panc-1. We isolated ALDH high , ALDH low and ALDH neg subsets according to the level of ALDH expression in the ALDH+ cells, and analyzed CD133/CD44 and the cell cycle. Results 1. The expression of ALDH in H22,LS-174T, SW-1990 and Panc-1was positive but negative in BGC-823 and SMMC-7721. 2. In H22 cell, ALDH high ，ALDH low and ALDH neg subsets were identified based on the difference of ALDH. 3. The proportion of CD133/CD44 in ALDH high ，ALDH low and ALDH neg subsets of H22 cell decreased. 4. During the observation of H22 cell, ALDH high subset existed in the G 0/G 1 phase, ALDH low subset was mainly in the S phase and ALDH neg subset largely appeared in the G 2/M phase. Conclusions There exists a specific subset between the stem cells and the normal cells in cancer cells, promising a new horizon in cancer treatment.
关键词
乙醛脱氢酶 /
肿瘤干细胞 /
流式细胞术 /
细胞表型 /
细胞周期
Key words
ALDH /
cancer stem cell /
flow cytometry /
cell phenotype /
cell cycle
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