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吴顺龙,李少林,李亚军.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[J].陆军军医大学学报(原第三军医大学学报),2016,38(22):2424-2430.
WU Shunlong,LI Shaolin,LI Yajun.Effects of 2-methoxyestradiol on proliferation, migration and radiosensitivity of nasopharyngeal carcinoma CNE-2 stem cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(22):2424-2430.
吴顺龙,李少林,李亚军.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[J].陆军军医大学学报(原第三军医大学学报),2016,38(22):2424-2430. DOI:
WU Shunlong,LI Shaolin,LI Yajun.Effects of 2-methoxyestradiol on proliferation, migration and radiosensitivity of nasopharyngeal carcinoma CNE-2 stem cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(22):2424-2430. DOI:
目的 探讨2-甲氧基雌二醇(2-methoxyestradiol,2-MeOE2)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-MeOE2浓度分为3组,分别用0、4、8 μmol/L 2-MeOE2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4 μmol/L 2-MeOE2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-MeOE2处理以及4 μmol/L 2-MeOE2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、8 μmol/L 2-MeOE2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(P0.01);迁移能力降低(P0.01);放疗敏感性增高,4 μmol/L 2-MeOE2 组放射增敏比(sensitivity enhancement ratio
SER)为1.19,8 μmol/L 2-MeOE2组SER为1.39;HIF-1α、NF-κB p65蛋白表达降低(P0.01),呈浓度依赖性。2-MeOE2 (4 μmol/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(P0.01)
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-MeOE2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=MeOE2抑制HIF=1α和NF-κB-p65蛋白表达。
Objective To investigate the effect and conceivable mechanism of 2-methoxyestradiol (2-MeOE2) on the proliferation
migration and 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 μmol/L). After the treatment
the cells were further irradiated by X-rays at the doses 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 (P0.01). Radiation-related parameters D0
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 (P0.01). 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 (P0.01). Conclusion Compared with CNE-2 cells
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 of expression of HIF-1α and NF-κB p65.
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