舌鳞癌Tca8113细胞株干细胞相关差异表达基因的筛选及分析

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[摘要] 目的 应用抑制消减杂交（SSH）技术筛选舌鳞癌干细胞相关差异表达基因。方法 以舌鳞癌tca8113细胞株中的醛脱氢酶（ALDH）高表达（ALDHbr）细胞与低表达（ALDHlow）细胞互为实验组和对照组，进行正反向消减杂交后建立差异表达基因文库，随机挑选差异基因克隆进行测序及生物信息学分析。结果 经Blast比对分析，得到62条差异表达基因；功能聚类发现多数与细胞周期调节、细胞分化等生物行为相关；信号通路分析显示有11条基因参与的12条信号通路发生显著改变。结论 应用SSH技术成功筛选出了与舌鳞癌干细胞相关的一些基因，进一步研究这些基因对探索舌鳞癌干细胞特异性表面标志物具有重要意义。

[关键词] 舌鳞状细胞癌； 肿瘤干细胞； 醛脱氢酶； 基因

[中图分类号] Q 786 [文献标志码] A [doi] 10.3969/j.issn.1000-1182.2012.05.003

Screen and analysis of differentially expressed genes related to stem-like cells in tongue squamous cell car-cinoma Tca8113 cell line Zou Bo, Ji Ping, Sun Shoujuan, Qi Xiaojuan. （Dept. of Oral and Maxillofacial Surgery, Hospital of Stomatology, Chongqing Medical University; Chongqing Research Center for Oral Diseases and Biome-dical Science, Chongqing 401147, China）

[Abstract] Objective To screen the differentially expressed genes of stem-like cells in tongue squamous cell car-cinoma using suppression subtractive hybridization（SSH）. Methods The cells with high and low expression of aldehyde dehydrogenase（ALDHbr and ALDHlow） were isolated respectively from human tongue squamous cell carcinoma Tca8113 cell line. SSH was used to identify differentially expressed genes in those two groups. Positive cell clones with high/

low-expressed ALDH were screened randomly for the genetic sequences and function analysis by the bioinformatics technology. Results Total 62 differentially expressed genes were identified, most of which got involved in regulation of cell cycle, cellular differentiation and other biology activities as gene ontology（GO） analysis indicated. Signal path-

way analysis revealed 12 pathways related to 11 genes changed significantly. Conclusion Several potential novel bio-marker candidates of tongue cancer stem cells were identified and they may provide important insights into the func-tions and characteristics of human tumor stem cell.

[Key words] tongue squamous cell carcinoma； tumor stem cell； aldehyde dehydrogenase； gene

肿瘤干细胞假说认为：肿瘤内存在一小部分细胞与肿瘤的发生、发展及转移密切相关，它们与正常组织干细胞具有相似的生物学特性，因而称为“肿瘤干细胞”[1-2]，目前已在多种肿瘤中证实了这群细

胞的存在[3-4]。肿瘤干细胞假说为解释肿瘤的发生机制提供了一个全新的理念，因而成为近几年研究的热点。

醛脱氢酶（aldehyde dehydrogenase，ALDH）是

一种多功能的蛋白，参与乙醛以及其他醛类的氧化，并与组胺及维甲酸的新陈代谢相关[5]。在肿瘤中ALDH常过表达，因为其能够降解细胞中的内源性和外源性有毒分子，导致肿瘤细胞生存期延长及化疗中耐药等问题的出现。最近的研究发现：ALDH还是一个能够有效富集肿瘤干细胞的标志物，在多种肿瘤中（特别是乳腺癌）分选出的高表达ALDH的肿瘤细胞都具有在免疫缺陷小鼠体内成瘤的能力，而且该亚群细胞还是一个独立的预后标志，表现出肿瘤干细胞的特性[6]。本实验采用荧光激活细胞分选（fluore-scence activated cell sorting，FACS）技术分选舌鳞癌

Tca8113细胞株中高表达ALDH（ALDHbr）的干细胞样细胞以及低表达ALDH（ALDHlow）的普通肿瘤细胞，利用抑制消减杂交（suppression subtractive hybridization，