CD133作为喉癌肿瘤起始细胞标志的实验研究  

Experimental investigation of CD133 as a 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,上海200031 [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》2006年第12期940-944,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的研究CD133在人喉癌细胞系Hep-2中的表达,观察纯化的CD133+肿瘤细胞体外生长特性,确定喉癌肿瘤起始细胞的表面标志。方法免疫荧光细胞化学技术及流式细胞仪检测喉癌Hep-2细胞系中的CD133表达,免疫磁珠分选技术纯化CD133+肿瘤细胞,体外培养并观察其增殖及分化能力。结果喉癌Hep-2细胞系中有3·22%的微量细胞CD133呈阳性表达,免疫磁珠富集的CD133+肿瘤细胞在无血清培养基中3、5、7d的吸光度分别为0.320、0.370、0.558,均高于相同条件下未分选细胞和CD133-细胞;CD133+在培养体系中的比例逐日下降,至培养的第12天,由第1天的90.88%下降至4.53%。结论喉癌Hep-2细胞系中,CD133+癌细胞有比其他细胞亚群强的体外分化和增殖能力,CD133是肿瘤起始细胞的标志之一。Objective To detect the expression of CD133 in human larynx tumor cell line, Hep-2 cell line and observe proliferation and differentiation ability of CD133^＋ groups in vitro. Methods Immunocytochemical staining and flow cytometry were used to detect the expression of putative tumorinitiating cell marker CD133 in Hep-2 cell line, and the selective technique of immunomagnetic beads was applied to purify CD133 positive cells. CD133^＋ tumor cells were cultured and their ability of proliferation and differentiation were observed in vitro. Results Only 3.22% of cells in Hep-2 cell line expressed CD133. In serum-free RPMI1640, On days 3, 5 and 7, their UV absorption were 0.320,0. 370 and 0.558 respectively. Compared with CD133^- cells and control Hep-2 cells, CD133 ^＋ cells demonstrated increased proliferation capacity. The proportion of CD133^＋ cells decreased in culture as days passed. In twelve days of culture, the percentage of CD133 ^＋ cells decreased from 90.88% to 4.53 %. Conclusions CD133 was one of makers for tumor-initiating cell of human laryngeal carcinoma, Hep-2 cell line. Its identification would provide a helpful tool to investigate the tumorigenic process of human laryngeal carcinoma and to develop targeted therapies.

关 键 词：喉肿瘤 肿瘤干细胞 肿瘤细胞 培养的 抗原 CD 

分 类 号：R739.65[医药卫生—肿瘤]