喉癌干细胞的培养及其放射抗拒相关miRNA的筛选研究  

Screening of radioresistance-related miRNA from laryngeal cancer stem cells

作　　者：段广亮[1] 戴辉萍 唐婷婷[1] 丁飞[1] 朱桂婷[1] 

机构地区：[1]杭州师范大学附属医院肿瘤诊治中心,310015

出　　处：《浙江医学》2016年第5期326-328,344,共4页Zhejiang Medical Journal

基　　金：2012年杭州市科技计划引导项目[杭科计(2012)259号(17)]

摘　　要：目的通过培养Hep-2喉癌细胞株,分离并扩增出肿瘤干细胞,放疗后筛选出调控放射抗拒的miRNA。方法含生长因子的无血清培养基培养Hep-2喉癌细胞株,流式细胞仪检测细胞中侧群细胞的比例。当侧群细胞比例达25%时,收集全部细胞球,筛选出CD133^+、CD44^+细胞作为喉癌干细胞,×线照射喉癌干细胞。miRNA生物芯片检测照射与未照射的喉癌干细胞,筛选出喉癌干细胞中2倍差异表达的miRNA,确定用以调控放射抗拒的miRNA。结果成功培养出侧群细胞,流式细胞仪检测其比例达到26.2%,筛选出的CD133^+、CD44^+细胞的比例均高于91%。通过miRNA芯片成功检测出喉癌干细胞中2倍差异表达的miRNA。结论本研究成功培养出喉癌干细胞,成功筛选出喉癌干细胞中2倍差异表达的miRNA,发现了可能用以调控放射抗拒的miRNA。Objective To screen radioresistance-related miRNA from laryngeal cancer stem cells. Methods Laryngeal cancer Hep-2 cells were cultured in SFM containing growth factor, and cancer stem cells （CD133^＋ CD44^＋） were detected and isolated with flow cytometry. The isolated laryngeal cancer stem cells were exposed to high energy X-ray. The differentially expressed miRNA （2-ford） were screened with miRNA biological chips in irradiated and non-irradiated laryngeal cancer stem celts. Results Overtop 91% CD133^＋ CD44^＋cells were harvested by FACS. The miRNAs of 2-fold differential expression were successfully screened from irradiated stem cells. Conclusion Laryngeal cancer stem cells have been isolated from Hep-2 cells, and miRNAs related to radioresistance have been screened from laryngeal cancer stem cells.

关 键 词：无血清培养基 MIRNA 侧群细胞 放疗 

分 类 号：R739.65[医药卫生—肿瘤]