CD133与喉癌肿瘤干细胞的实验研究  

Investigation of CD133 as putative marker of tumor-initiating cell in laryngeal carcinoma

作　　者：卫旭东[1] 周梁[1] 程磊[1] 田洁[2] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻咽喉科 [2]复旦大学附属眼耳鼻喉科医院中心实验室,上海200031

出　　处：《中华耳鼻咽喉头颈外科杂志》2007年第9期692-696,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的探讨 CD133在人喉癌细胞系 Hep-2中的表达,观察纯化的 CD133阳性肿瘤细胞、CD133阴性肿瘤细胞及未分选 Hep-2细胞在重症联合免疫缺陷小鼠中的成瘤性,筛选 Hep-2细胞系肿瘤干细胞的表型。方法流式细胞仪检测 CD133在 Hep-2细胞系中的表达,免疫磁珠分选技术纯化 CD133阳性肿瘤细胞,分选所得各细胞亚群细胞以及未分选细胞以一定的数量级注入重症联合免疫缺陷小鼠腹部皮下,比较其成瘤差异性。分选后的细胞进行了细胞周期的分析和 HE 染色观察生长形态,以排除成瘤差异由细胞周期分布不同及异源细胞引起。结果流式细胞仪示 CD133在Hep-2细胞系中呈微量恒定表达,表达概率(3.15±0.83)%。免疫磁珠富集的 CD133阳性肿瘤细胞并不处于细胞周期生长旺盛时相或优势分裂时相,而是与分选前周期分布相似且均匀的一类细胞;HE 染色示细胞形态亦符合恶性肿瘤细胞的病理生长特性。体外成瘤试验显示16/20个 CD133阳性细胞注射部位、7/20个 CD133阴性细胞注射部位、10/20个未分选细胞注射部位可见肿瘤生长,统计学分析表明 CD133阳性肿瘤细胞较 CD133阴性细胞(X^2=8.286,P=0.004)、未分选细胞(X^2=3.956,P=0.047)在重症联合免疫缺陷小鼠体内具有很强的成瘤性。结论喉癌 Hep-2细胞系中,CD133阳性癌细胞具有强的体内增殖能力,CD133为喉癌肿瘤干细胞的标志之一。Objective To detect the expression of CD133 in human established larynx tumors cell line,Hep-2 cell line and observe tumorigenicity of CD133 （ ＋ ） tumorigenic cells, CD133 （ - ） tumorigenic cells and unsorted Hep-2 cells in vivo. The marker of cancer stem cells in Hep-2 cell line was explored. Methods Flow cytometry was used to detect the expression of putative tumor-initiating cell marker CD133 in Hep-2 cell line. The immunomagnetic beads separation was applied to purify CD133 positive cells. CD133 （ ＋ ） tumor cells , CD133 （ - ） tumor cells and unsorted Hep-2 cells were injected to severe combined immune deficiency mice individually to observe their ability of forming new tumors. To determine whether the difference in tumorigenicity in subpopulation of Hep-2 cells was due to differences in cell growth activity and cell cycle. The growth of sorted cells in vitro was observed with HE stain and analyzed cell cycle of CD133 （ ＋ ） cells and unsorted cells by flow cytometry. Results Only a small proportion （3. 15 ± 0. 83 ） % of cells in Hep-2 cell line express CD133. Both unsorted cells and sorted cells were all consistent with the character of malignant tumor. Comparison of the cell cycle status of sorted and unsorted cancer cells after magnetic sorting revealed that both cells exhibited a similar cell cycle distribution. In 20 injection sites, 16 sites contained tumor in CD133（ ＋ ） group ,whereas only 10 sites in unsorted group, 7 sites in CD133（ - ） group contained tumor. Compared with CD133 （ - ） group （ χ^2 = 8.286, P = 0. 004） and unsorted group （χ^2 = 3. 956, P = 0. 047 ）, CD133 positive cells possessed a marked capacity for giving rise to new tumors in vivo. Conclusions CD133 is one of makers for cancer stem cell in human larynx tumors, Hep-2 cell line. Identification of it provides a powerful tool to investigate the tumorigenic process in the larynx and to develoo theraoies targeted to the tumor-initiating cell.

关 键 词：喉肿瘤 肿瘤干细胞 细胞系 

分 类 号：R739.65[医药卫生—肿瘤]