人鼻咽癌细胞株分选CD133^+细胞实验研究

Sorting of CD133^+ positive cells from nasopharyngeal carcinoma cell line

目的观察CD133在人鼻咽癌细胞株中的表达,探讨CD133+肿瘤细胞的体外增殖及分化能力从而确定鼻咽癌肿瘤干细胞的标志。方法使用免疫细胞化学及流式细胞技术检测鼻咽癌CNE2Z细胞株中的表达,免疫磁珠分选技术纯化肿瘤细胞,体外培养,观察其增殖及分化能力。结果鼻咽癌CNE2Z细胞株中有0.168%呈微量阳性表达,利用免疫磁珠分选技术纯化细胞的百分比为87.2%,免疫磁珠富集的CD133+肿瘤细胞在无血清培养基中1、3、5、7天的吸光度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87.2%下降至0.1%。结论鼻咽癌CNE2Z细胞株中,CD133+癌细胞有比其他细胞亚群强的体外分化和增殖能力,具有自我更新、生成其他表型肿瘤细胞等干细胞样特性,CD133可能是肿瘤起始细胞的标志之一。

Purpose To detect the expression of CD133 in human nasopharyngeal carcinoma cell line, CNE2Z cell line and to observe the proliferation and differentiation ability of CD133＋ groups in vitro. Methods Immunocytochemical staining and flow eytometry were used to detect the expression of putative tumor initiating cell marker CD133 in CNE2Z cell line, and the selective technique of inmunomagnetie 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 0. 168% of cells in CNE2Z cell line expressed CD133. In senun-ffee RPMI1640, on days 1, 3, 5 and 7, their UV absorption was higher in CD133 cells than control CNE2Z cells. CD133＋ cells demonstrated increased proliferating capacity. The proportion of CD133＋ cells decreased in culture as days passed. In twelve days of culture, the percentage of CD133＋ cells decreased from 87.2% to 0. 1%. Conclusions CD133＋ cells process stronger motility than CD133 - , CNE2Z cells in vitro and CD133＋ cells may contribute to the metastsis of nasopharyngeal carcinoma cell. Therefore, CD133 may be one of makers for tumor-initiating cell in human nasopharyngeal carcinoma, CNE2Z cell line.