[1]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].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 Third Mil Med Univ,2016,38(22):2424-2430.
点击复制

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(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第22期
页码:
2424-2430
栏目:
基础医学
出版日期:
2016-11-30

文章信息/Info

Title:
Effects of 2-methoxyestradiol on proliferation, migration and radiosensitivity of nasopharyngeal carcinoma CNE-2 stem cells
作者:
吴顺龙李少林李亚军
重庆医科大学附属第一医院肿瘤科;重庆医科大学基础医学院放射医学与肿瘤教研室;遵义医学院附属医院肿瘤科; 遵义医学院第三附属医院肿瘤科
Author(s):
WU Shunlong LI Shaolin  LI Yajun

Department of Oncology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016;  Department of Radiation Oncology, College of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016; Department of Oncology, Affiliated Hospital of Zunyi Medical College, Zunyi,  563000, Guizhou Province;  Department of Oncology, Third Affiliated Hospital of Zunyi Medical College, Zunyi, 563000, Guizhou Province, China

关键词:
鼻咽癌癌干细胞HIF-1&alpha2-MeOE2NF-&kappaB-p65放疗敏感性
Keywords:
nasopharyngeal carcinomacancer stem cellsHIF-1&alpha2-MeOE2NF-&kappaB p65radiosensitivity
分类号:
R739.63;R965;R977.12
文献标志码:
A
摘要:

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(P<0.01);迁移能力降低(P<0.01);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(P<0.01),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(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-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。

Abstract:

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 (P<0.01). 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 (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 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.

参考文献/References:

[1]王志海, 曾泉, 陈弢, 等. 抑制NFBD1基因的表达对鼻咽癌CNE-1细胞放射敏感性的影响[J]. 肿瘤, 2014, 34(2): 101-107. DOI: 10.3781/j.issn.1000-7431
[2]Long H, Xie R, Xiang T, et al. Autocrine CCL5 signaling promotes invasion and migration of CD133+ ovarian cancer stem-like cells via NF-κB-mediated MMP-9 upregulation[J]. Stem Cells, 2012, 30(10): 2309-2319. DOI: 10.1002/stem.1194
[3]Yu S, Zhang R, Liu F, et al. Notch inhibition suppresses nasopharyngeal carcinoma by depleting cancer stem-like side population cells[J]. Oncology Reports, 2012, 28(2): 561-566.DOI: 10.3892/or.2012.1830
[4]Guo D, Xu B L, Zhang X H, et al. Cancer stem-like side population cells in the human nasopharyngeal carcinoma cell line cne-2 possess epithelial mesenchymal transition properties in association with metastasis[J]. Oncology Reports, 2012, 28(1): 241-247.DOI: 10.3892/or.2012.1781
[5]Oliveira L R, Oliveira-Costa J P, Soave D F, et al. P177. Cancer stem cell immunophenotypes in oral squamous cell carcinoma[J]. Oral Pathol Med, 2011, 40(2): 135-142.DOI: 10.1111/j.16000714.2010.00967
[6]Annunziata C M, Stavnes H T, Kleinberg L, et al. Nuclear Factor kappa B Transcription Factors Are Coexpressed and Convey a Poor Outcome in Ovarian Cancer[J]. Cancer, 2010, 116(13): 3276-3284.DOI: 10.1002/cncr.25190
[7]Masi C M, Hawkley L C, Cacioppo J T. Serum 2-methoxyestradiol, an estrogen metabolite, is positively associated with serum HDL-C in a populationbased sample[J]. Lipids, 2012, 47(1): 35-38.DOI: 10.1002/ cncr.25190
[8]Qi B, Chen H L, Shang D, et al. Effects of hypoxia-inducible factor-1α and matrix metalloproteinase-9 on alveolarcapillary barrier disruption and lung edema in rat models of severe acute pancreatitis-associated lung injury[J]. Experimental & Therapeutic Med, 2014, 8(3): 899-906.DOI: 10.3892/etm.2014.1810
[9]Xie H C, He J P, Zhu J F, et al. Expression of HIF-1alpha and VEGF in skeletal muscle of plateau animals in response to hypoxic stress[J]. Physiological Res, 2014, 63(6): 801-805.
[10]Pan H, Chen S, Yuan W, et al. Down-regulated CBS/H2S pathway is involved in high-salt-induced hypertension in Dahl rats[J]. Nitric Oxide Biology & Chemistry, 2015, 46: 192-203.DOI: 10.1016/j.niox.2015.01.004
[11]Ricker J L, Chen Z, Yang X P, et al. 2-Methoxyestradiol inhibits hypoxiainducible factor 1α, tumor growth, and angiogenesis and augments paclitaxel efficacy in head and neck squamous cell carcinoma[J]. Clin Cancer Res, 2004, 10(24): 8665-8673.DOI: 10.1158/1078-0432.CCR-04-1393
[12]Parrondo R, Pozas D L A, Reiner T, et al. NF-κB activation enhances cell death by antimitotic drugs in human prostate cancer cells[J]. Molecular Cancer, 2010, 9(2): 1-13.DOI:  10.1186/1476-4598-9-182
[13]Kumar A P, Garcia G E, Orsborn J, et al. 2-Methoxyestradiol interferes with NFκB transcriptional activity in primitive neuroectodermal brain tumors: implications for management [J]. Carcinogenesis, 2003, 24(2): 209-216.DOI: 10.1093/carcin/24.2.209
[14]Zhang X Y, Zhan R, Huang H B, et al. Mechanism underlying 2-methoxyestradiol inducing apoptosis of K562 cells [J]. Zhongguo shi yan xue ye xue za zhi, 2009, 17(2): 340-344.
[15]Sutherland T E, Anderson R L, Hughes R A, et al. 2-Methoxyestradiol—a unique blend of activities generating a new class of anti-tumour/anti-inflammatory agents[J]. Drug Discov Today, 2007, 12: 577-584.DOI: 10.1016/ j.drudis.2007.05.005
[16]Zhao C, Setrerrahmane S, Xu H. Enrichment and characterization of cancer stem cells from a human nonsmall cell lung cancer cell line[J]. Oncology Reports, 2015.DOI: 10.3892/or.2015.4163
[17]MirandaLorenzo I, Dorado J, Lonardo E, et al. Intracellular autofluorescence: a biomarker for epithelial cancer stem cells[J]. Nature Methods, 2014, 11(11): 1161-1169. DOI: 10.1038/nmeth.3112
[18]Li Y J, Wu S L, Lu S M, et al. (-)-Epigallocatechin-3-gallate inhibits nasopharyngeal cancer stem cell selfrenewal and migration and reverses the epithelial-mesenchymal transition via NF-κB p65 inactivation[J]. Tumor Biology, 2015, 36(4): 2747-2761.DOI: 10.1007/s13277-014-2899-4
[19]李亚军, 郭颖, 吴顺龙, 等. 人鼻咽癌干细胞微球体培养并鉴定其生物特性[J]. 医学研究生学报, 2014, 27(11): 1133-1138.DOI: 10.3969/j.issn.1008-8199.2014.11.004
[20]Reumann S, Shogren K L, Yaszemski M J, et al. Inhibition of Autophagy Increases 2-MethoxyestradiolInduced Cytotoxicity in SW1353 Chondrosarcoma Cells[J]. J Cell Biochem, 2016, 117(3): 751-759. DOI: 10.1002/jcb.25360
[21]Shigdar S, Qiao L, Zhou S F, et al. RNA aptamers targeting cancer stem cell marker CD133[J]. Cancer letters, 2013, 330(1): 84-95.DOI: 10.1016/j.canlet.2012.11.032
[22]Zhang X, de Milito A, Olofsson M H, et al.Targeting Mitochondrial Function to Treat Quiescent Tumor Cells in Solid Tumors[J]. Mol Sci, 2015, 16(11): 27313-27326.DOI: 10.3390/ijms161126020
[23]Yang P, Reece E A. 45: Role of HIF-1alpha in maternal hyperglycemia-induced embryonic vasculopathy[J]. American Journal of Obstetrics & Gynecology, 2011, 204(1): S27. DOI: 10.1016/j.ajog.2011.01.012
[24]Tang N, Wang L, Esko J, et al. Loss of HIF-1α in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis[J]. Cancer Cell, 2004, 6(5): 485-495. DOI: 10.1016/j.ccr.2004.09.026
[25]Li H, Chen J, Zen W, et al.Effect of hypoxia inducible factor-1 antisense oligonucleotide on liver cancer [J].Int Clin Exp Med,2015,8(8): 12650-12655.
[26]Zhang H, Qian D Z, Tan Y S, et al. Digoxin and other cardiac glycosides inhibit HIF-1α synthesis and block tumor growth[J]. Proceedings of the National Academy of Sciences, 2008, 105(50): 19579-19586.DOI: 10.1073/pnas.0809763105
[27]Didonato J A, Frank M, Michael K. NF-κB and the link between inflammation and cancer[J]. Immunological Reviews, 2012, 246(1): 379-400.DOI: 10.1111/j.1600065X.2012. 01099.x
[28]Luqman S, Pezzuto J M. NF-κB: a promising target for natural products in cancer chemoprevention[J]. Phytotherapy Res, 2010, 24(7): 949-963.DOI: 10.1002/ptr.3171
 [29]Chen C, Yu Z. siRNA targeting HIF-1alpha induces apoptosis of pancreatic cancer cells through NFkappaB-independent and -dependent pathways under hypoxic conditions[J]. Anticancer Res, 2009, 29(4): 1367-1372.
[30]Rius J, Guma M, Schachtrup C,et al. NF-κB links innate immunity to the hypoxic response through transcriptional regulation of HIF-1α[J]. Nature,2008,453: 807-811.DOI: 10.1038/nature06905
[31]Belaiba R S, Bonello S, Zhringer C, et al. Hypoxia up-regulates hypoxia-inducible factor-1α transcription by involving phosphatidylinositol 3-kinase and nuclear factor κB in pulmonary artery smooth muscle cells [J]. Mol Biol Cell, 2007, 18: 4691-4697.DOI: 10.1091/mbc.E07-04-0391

相似文献/References:

[1]金世龙,黄中荣,陈华,等.HuH7细胞系CD13+CD133+HCC细胞分选及CSCs特性分析[J].第三军医大学学报,2012,34(16):1658.
 Jin Shilong,Huang Zhongrong,Chen Hua,et al.Sorting and cancer stem cell characteristic analysis of CD13+CD133+HCC cells in HuH7 cell line[J].J Third Mil Med Univ,2012,34(22):1658.
[2]金世龙,余天雾,黄中荣,等.As2O3在HuH7及CD13+CD133+LCSCs增殖和分化中作用的初步观察[J].第三军医大学学报,2012,34(18):1857.
 Jin Shilong,Yu Tianwu,Huang Zhongrong,et al.Role of arsenic trioxide in proliferation of HuH7 cells and differentiation of CD13+CD133+ LCSCs[J].J Third Mil Med Univ,2012,34(22):1857.

更新日期/Last Update: 2016-11-23