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目的:从人鼻咽癌SUNE-1 5-8F细胞株中分离出CD44+细胞,并对其放化疗敏感性进行初步研究。方法:常规培养SUNE-1 5-8F细胞,采用免疫荧光技术及流式细胞学技术检测SUNE-1 5-8F细胞中CD44+的表达并用流式细胞仪分选CD44+细胞;采用四甲基偶氮唑蓝(MTT)法、克隆形成实验等检测并比较CD44+、CD44-细胞在放化疗敏感性上的差异。结果:CD44+细胞在鼻咽癌细胞中SUNE-1株所占的比率约为52.5%;新分选的CD44+和CD44-细胞在接受2Gy放射处理后,其平均克隆生成效率分别为(23.44±1.90)%和(7.78±1.17)%(P<0.001);CD44+细胞较CD44-细胞在相同顺铂和多西他赛药物浓度下显示出更高的细胞存活率。结论:流式细胞学技术能够有效地分选细胞;CD44+细胞较CD44-具有更强的放化疗抗拒性,CD44很有可能成为鼻咽癌肿瘤干细胞分子标记物之一。
OBJECTIVE: To isolate CD44 + cells from human nasopharyngeal carcinoma SUNE-1 5-8F cell line and to study the radiosensitivity and chemosensitivity. Methods: SUNE-1 5-8F cells were cultured routinely. The expression of CD44 + in SUNE-1 5-8F cells was detected by immunofluorescence and flow cytometry. CD44 + cells were sorted by flow cytometry. MTT assay and clonogenic assay were used to detect and compare the chemosensitivity of CD44 + and CD44- cells. Results: The percentage of CD44 + cells in nasopharyngeal carcinoma cells was 52.5%. The average clonogenic efficiency of newly sorted CD44 + and CD44- cells was (23.44 ± 1.90 ) And (7.78 ± 1.17)%, respectively (P <0.001); CD44 + cells showed higher cell survival rates than CD44- cells at the same cisplatin and docetaxel drug concentrations. Conclusion: Flow cytometry can effectively differentiate cells; CD44 + cells have stronger chemoradiotherapy and chemosensitivity than CD44-, and CD44 is likely to be one of the molecular markers of NPC stem cells.