【摘要】 背景:在多种肿瘤中已证实CD34与肿瘤的发生、发展、复发、转移、耐药密切相关,但是CD34与鼻咽癌的关系目前还知之甚少。目的:分选鼻咽癌细胞株中CD34+与CD34-细胞的增殖和迁移能力。方法:利用流式细胞术检测CD34在鼻咽癌细胞株5-8F、6-10B、CNE1、CNE2中的表达,根据细胞表面标志物分选出CD34+与CD34-细胞,进行纯度鉴定;通过体外MTT法、平板克隆形成实验以及划痕实验分析CD34+细胞与CD34-细胞增殖和迁移能力。结果与结论:(1)4种鼻咽癌细胞株中CD34表达均恒定在0.1%-0.2%,且CD34水平与细胞的生长密度关系密切;(2)分选的CD34+细胞群中,CD34+细胞例能达到98%以上,在CD34-细胞群中未能检测到CD34+细胞;(3)CD34+细胞在第1,3,5,7天增殖速度显著高于CD34-细胞(P<0.05);(4)CD34+细胞克隆形成率显著高于CD34-细胞(P<0.05);(5)CD34+细胞向划痕区迁移的细胞明显多于CD34-细胞(P<0.05);(6)结果提示,CD34+细胞体外培养具有更高的增殖和转移能力,具有鼻咽癌肿瘤干细胞潜能。更多还原

【Abstract】 BACKGROUND: Previous research have confirmed that CD34 is closely related to oncogenesis, progress, recurrence, metastasis and drug-resistance of various cancers, but its role in nasopharyngeal carcinoma remains unclear. OBJECTIVE: To sort cells positive and negative for CD34 in nasopharyngeal carcinoma cell lines and to detect cell proliferation and migration. METHODS: Expressions of CD34 in nasopharyngeal carcinoma cell lines 5-8F, 6-10 B, CNE1 and CNE2 were detected by flow cytometry. And CD34+ and CD34- cells were sorted based on cell surface markers for purity identification. Afterwards, proliferation and migration of CD34+ and CD34- cells were detected by MTT assay, colony-formation assay and scratch assay. RESULTS AND CONCLUSION: All four nasopharyngeal carcinoma cell lines expressed CD34 in 0.1%-0.2%, and the level of CD34 was closely related to the cell growth density. The purity of CD34+ cell was more than 98% in the sorted CD34+ cell populations, but no CD34+ cells were found in the sorted CD34- cell populations. At 1, 3, 5 and 7 days the proliferation rate of CD34+ cell, populations was significantly higher than that of CD34- cells(P < 0.05). Consistently, the colony-formation efficiency of CD34+ cell was significantly higher than that of CD34- cells(P < 0.05). Moreover, CD34+ cells migrated significantly faster than CD34- cells by scratch assay(P < 0.05). In conclusion, CD34+ cells cultured in vitro display higher proliferation and migration capacities, indicating that CD34+ cells have the potential of nasopharyngeal carcinoma stem cells.更多还原