CD44^+鼻咽癌细胞放化疗敏感性研究

Sensitivity of CD44^+ Nasopharyngeal Carcinoma Cells to Chemoradiotherapy

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 study the biological features of CD44＋ cells separated from the human nasopharyngeal carcinoma（NPC） SUNE-1 5-8F cell line.Methods： Immunocytochemistry and flow cytometry were used to detect the expression of CD44 in SUNE-1 5-8F.Fluorescence-Activated Cell Sorting（FACS） was applied to purify CD44＋ cells.MTT assay or Clone formation assay were used to detect the differences of CD44＋ and CD44-cells in radiosensitivty and chemosensitivity in vitro.Results： CD44 was positively expressed in about 52.5% of NPC SUNE-1 5-8F cells.After 2 Gy radiation,the average clone formation efficiency for CD44＋ and CD44-cells were respectively（23.44±1.90）% and（7.78±1.17） %（P0.001）.After Cisplatin and Docetaxel treatment,CD44＋ cells showed a higher survival rate in the same drug concentration compared with CD44-cells.Conclusion： FACS isolation of cells is an effective method.CD44＋ cells with strong resistance to chemotherapy drugs and X-rays may be assumed as one of markers for treatment of NPC tumor stem cells.