2-甲氧基雌二醇对鼻咽癌CNE-2干细胞体外增殖、迁移及放疗敏感性的影响  

Effects of 2-methoxyestradiol on proliferation,migration and radiosensitivity of nasopharyngeal carcinoma CNE-2 stem cells

作　　者：吴顺龙[1] 李少林[2] 李亚军[3,4] 

机构地区：[1]重庆医科大学附属第一医院肿瘤科,重庆400016 [2]重庆医科大学基础医学院放射医学与肿瘤教研室,重庆400016 [3]遵义医学院附属医院肿瘤科,贵州遵义563000 [4]遵义医学院第三附属医院肿瘤科,贵州遵义563000

出　　处：《第三军医大学学报》2016年第22期2424-2430,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金(81560444);贵州省卫计委立项课题(gzwjkj2014-2-167);贵州省科技厅社会发展攻关课题(黔科合SY字[2015]3046号)--子课题~~

摘　　要：目的探讨2-甲氧基雌二醇(2-methoxyestradiol,2-Me OE2)对人鼻咽癌CNE-2干细胞(nasopharyngeal cancer stem cells,NPCSCs)增殖、迁移及放疗敏感性的影响及其可能机制。方法收集本课题组前期富集并已鉴定的鼻咽癌CNE-2干细胞,Western blot检测亲本CNE-2细胞及NPCSCs乏氧诱导因子-1α(HIF-1α)和核转录因子(NF-κB p65)的蛋白表达,克隆形成实验检测两组细胞的放射敏感性。按2-Me OE2浓度分为3组,分别用0、4、8μmol/L 2-Me OE2处理鼻咽癌干细胞,倒置显微镜观察细胞形态变化,CCK-8及Transwell检测细胞增殖及迁移能力,再联合X射线(0、2、4、8 Gy)照射,克隆形成实验检测3组细胞放疗敏感性;同时用4μmol/L 2-Me OE2处理鼻咽癌干细胞后不同时间(0、24、48 h)提取蛋白,Western blot检测不同浓度2-Me OE2处理以及4μmol/L 2-Me OE2处理后0、24、48 h提取的干细胞HIF-1α和NF-κB p65蛋白表达。结果相对于亲本CNE-2细胞,干细胞HIF-1α和NF-κB p65蛋白表达明显增高(P均<0.01),X射线辐射后干细胞各放射学参数D0、Dq、N及SF2均高于亲本CNE-2细胞。0、4、8μmol/L 2-Me OE2处理干细胞后,随浓度增加,致密球状生长的干细胞微球体逐渐变得离散,干细胞增殖活力降低(P<0.01);迁移能力降低(P<0.01);放疗敏感性增高,4μmol/L2-Me OE2组放射增敏比(sensitivity enhancement ratio,SER)为1.19,8μmol/L 2-Me OE2组SER为1.39;HIF-1α、NF-κB p65蛋白表达降低(P<0.01),呈浓度依赖性。2-Me OE2(4μmol/L)处理干细胞后随时间延长HIF-1α、NF-κB p65蛋白表达明显降低(P<0.01),呈时间依赖性。结论相对于亲本CNE-2细胞,干细胞高表达HIF-1α和NF-κB p65,并且对放疗具有较低的敏感性。2-Me OE2能有效抑制鼻咽癌CNE-2干细胞增殖、迁移并增加其放疗敏感性,其机制可能为2-Me OE2抑制HIF-1α和NF-κB p65蛋白表达。（2-MeOE2） on the Objective To investigate proliferation, migration and the effect and conceivable mechanism of 2-methoxyestradiol radiosensitivity of nasopharyngeal carcinoma （NPC） CNE-2 stem cells in vitro. Methods Pre-identified and collected NPC CNE-2 stem cells from our group were used for this study. Nasopharyngeal carcinoma stem cells （NPCSCs） were treated by 2-MeOE2 at different concentrations （0, 4 and 8 μ mol/L）. After the treatment, the cells were further irradiated by X-rays at thedoses of 0, 2, 4 and 8 Gy. Total proteins were extracted in 0, 24 and 48 h after the cells treated by 2-MeOE2 （4 μmol/L）. Western blotting and colony formation assay were used to detect protein expression of HIF-1α and NF-κB p65 and radiosensitivity of CNE-2 and NPCSC. CCK-8 assay and transwell assay were used to measure the proliferation and migration of NPCSC. Results Western blot results showed that HIF-1α and NF-κB p65 were increasingly expressed in the NPCSCs compared with the parental CNE-2 cells （P 〈 0. 01 ）. Radiation-related parameters DO, Dq, N and SF2 had higher values in, NPCSCs than in CNE-2 cells. After treatment, the proliferation and migration capabilities of NPCSCs were decreased significantly along with increased 2-MeOE2 concentration （P 〈 0. 01 ）. 2-MeOE2 treatment also increased radiosensitivity in NPCSCs, showing sensitivity enhancment ratio （SER） values of 1. 19 and 1.39 at 4 and 8 μmol/L of 2-MeOE2, respectively. Moreover, the protein expression of HIF-1α and NF-κB p65 was decreased significantly along increased concentration and delayed harvested time, suggesting their expression patterns were concentration- and time-dependent （ P 〈 0. 01 ）. Conclusion Compared with CNE-2 ceils, NPCSCs have higher protein expression of HIF-1α and NF-κB p65, and are resist to radiotherapy. 2-MeOE2 could effectively reduce the proliferation and migration, and sensitize the radioresistance to NPC CNE-2 stem cells, which might mechanistically relate to 2-MeOE2 inhibition

关 键 词：鼻咽癌 癌干细胞 HIF-1α 2-MeOE2 NF-κB p65 放疗敏感性 

分 类 号：R739.63[医药卫生—肿瘤] R965[医药卫生—临床医学]