山东大学耳鼻喉眼学报 ›› 2017, Vol. 31 ›› Issue (5): 79-84.doi: 10.6040/j.issn.1673-3770.0.2017.321

• 论著 • 上一篇    下一篇

MiR-150调控Nanog对鼻咽癌侧群细胞增殖、侵袭的影响

周毅波,龚小蓉,于锋   

  1. 广州市第十二人民医院 广州市耳鼻咽喉头颈外科医院 广州医科大学耳鼻咽喉头颈外科研究所, 广东 广州 510620
  • 收稿日期:2017-07-26 出版日期:2017-10-16 发布日期:2017-10-16
  • 通讯作者: 于锋. E-mail:fishwoo@sina.com
  • 基金资助:
    广东省医学科学技术研究基金项目(A2015235);广州市医药卫生科技项目(20141A011045)

Effects of miR-150 targeted to Nanog on proliferation and invasion of nasopharyngeal carcinoma side population cells.

ZHOU Yibo, GONG Xiaorong, YU Feng   

  1. Guangzhou 12th Peoples Hospital, Guangzhou Hospital of Otolaryngology Head and Neck Surgery, Institute of Otolaryngology Head and Neck Surgery of Guangzhou Medical University, Guangzhou 510620, Guangdong, China
  • Received:2017-07-26 Online:2017-10-16 Published:2017-10-16

HTML

PDF (PC)

249

摘要： 目的 检测MiR-150在鼻咽癌侧群(SP)细胞中的表达,并探讨其是否通过调控靶基因Nanog促进鼻咽癌侧群细胞的增殖与侵袭。 方法 采用SYBR Green 实时定量荧光聚合酶链式反应(qRT-PCR)法检测MiR-150及Nanog在鼻咽癌侧群细胞和非侧群(NSP)细胞中的表达情况;并通过瞬时转染miR-150 inhibitor 至SP细胞、miR-150 mimic至NSP细胞中,western blotting检测Nanog的表达情况;进一步通过CCK-8实验、Transwall实验检测鼻咽癌侧群细胞增殖、侵袭能力的变化,数据处理采用两独立样本t检验。 结果 qRT-PCR检测结果表明,MiR-150在SP细胞(0.99±0.05)较NSP细胞(0.59±0.02)中表达水平升高(t=8.06, P<0.000 1),Nanog在鼻咽癌SP细胞(0.99±0.47)较NSP 细胞(0.49±0.05)表达升高(t=7.5, P<0.000 1);SP细胞转染MiR-150 inhibitor(0.46±0.03)较对照组(1.01±0.07)Nanog表达下调(t=6.85, P=0.000 5)、CCK-8实验示增殖受抑制、Transwell侵袭实验示侵袭能力减弱(114.40±5.14 vs 57.30±4.29, t=8.5, P<0.000 1);NSP细胞转染MiR-150 mimic(1.01±0.06)组较对照组(0.48±0.04)Nanog表达上调(t=6.16, P=0.000 8),增殖及侵袭能力增强。 结论 鼻咽癌SP细胞中MiR-150与Nanog呈高表达,MiR-150可通过调控靶基因Nanog促进鼻咽癌侧群细胞的增殖与侵袭。

关键词: SP细胞, 增殖, 侵袭, 鼻咽癌, MiR-150, NSP细胞

Abstract: Objective To detect the level of miR-150 in side population(SP)and non-side population(NSP)cells of nasopharyngeal carcinoma and to investigate whether miR-150 promoted proliferation and invasion via the Nanog gene. Method miR-150 and Nanog were detected by qRT-PCR in two cell types. An miR-150 inhibitor and miR-150 mimic were used to reduce and upregulate the level of miR-150, respectively, and CCK-8 and Transwell assays were performed to detect cell proliferation and invasion; the change in Nanog was measured by qRT-PCR and western blot analysis. A t-test for independent samples was used to analyze the data. Results The miR-150 level in SP cells(0.99±0.05)was significantly higher than that in NSP cells(0.59±0.02, t=8.06, P<0.000 1)and the level of Nanog in SP cells(0.99±0.47)was significantly higher than that in NSP cells(0.49±0.05, t=7.5, P<0.000 1). The CCK-8 and Transwell assays showed that MiR-150 inhibited proliferation and invasion(114.40±5.14 vs 57.30±4.29, t=8.5, P<0.000 1); western blotting showed that the reduction in MiR-150 decreased the protein expression of Nanog in SP cells(1.01±0.07 vs 0.46±0.03, t=6.85, P=0.000 5). In contrast, the miR-150 mimic group showed significant upregulation of Nanog in NSP cells(0.48±0.04 vs 1.01±0.06, t=6.16, P=0.000 8). Conclusion miR-150 and Nanog were highly expressed in SP cells of nasopharyngeal carcinoma; additionally, miR-150 promoted proliferation and invasion through upregulation of the Nanog gene in nasopharyngeal carcinoma.

Key words: MiR-150, Side population cells, Invasion, Non- side population cells, Proliferation, Nasopharyngeal carcinoma

中图分类号: 

  • R739.63
[1] Boumahdi S, Driessens G, Lapouge G, et al. SOX2 controls tumour initiation and cancer stem-cell functions in squamous-cell carcinoma[J]. Nature, 2014, 511(7508):246-250.
[2] Zhang HB, Ren CP, Yang XY, et al. Identification of label-retaining cells in nasopharyngeal epithelia and nasopharyngeal carcinoma tissues[J]. Histochem Cell Biol, 2007, 127(3):347-354.
[3] Wang J, Guo LP, Chen LZ, et al. Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoma cell line[J]. Cancer Res, 2007, 67(8):3716-3724.
[4] 刘付梅,李祥勇,周克元. miR-150与临床疾病研究的新进展[J].医学研究生学报,2014,27(2):194-198. LIU Fumei, LI Xiangyong, ZHOU Keyuan. New advances of miR-150 and diseases[J]. J Med Postgraduates, 2014, 27(2):194-198.
[5] Zhang J, Luo N, Luo Y, et al. MicroRNA-150 inhibits human CD133-positive liver cancer stem cells through negative regulation of the transcription factor c-Myb[J]. Int J Oncol, 2012, 40(3):747-756.
[6] Wu Q, Jin H, Yang Z, et al. MiR-150 promotes gastric cancer proliferation by negatively regulating the pro-apoptotic gene EGR2[J]. Biochem Biophys Res Commun, 2010, 392(3):340-345.
[7] Cao M, Hou D, Liang H, et al. miR-150 promotes the proliferation and migration of lung cancer cells by targeting SRC kinase signalling inhibitor 1[J]. Eur J Cancer, 2014, 50(5):1013-1024.
[8] Yin QW, Sun XF, Yang GT, et al. Increased expression of microRNA-150 is associated with poor prognosis in non-small cell lung cancer[J]. Int J Clin Exp Pathol, 2015, 8(1):842-846.
[9] 张先锋,黄远见,肖娟. miR-150促进鼻咽癌细胞增殖和侵袭的机制研究[J].中国全科医学,2016,19(21):2513-2517. ZHANG Xianfeng, HUANG Yuanjian, XIAO Juan. Mechanism research of miR-150 promoting the proliferation and invasion of nasopharyngeal carcinoma[J]. Chin Gen Prac, 2106, 19(21):2513-2517.
[10] Boyer LA, Lee TI, Cole MF, et al. Core transcriptional regulatory circuitry in human embryonic stem cells[J]. Cell, 2005, 122(6):947-956.
[11] Suzuki A, Raya A, Kawakami Y, et al. Maintenance of embryonic stem cell pluripotency by Nanog-mediated reversal of mesoderm specification[J]. Nat Clin Pract Cardiovasc Med, 2006, 3:S114-122.
[12] Yang L, Zhang X, Zhang M, et al. Increased Nanog expression promotes tumor development and Cisplatin resistance in human esophageal cancer cells[J]. Cell Physiol Biochem, 2012, 30(4):943-952.
[13] Jeter CR, Yang T, Wang JC, et al. NANOG in cancer stem cells and tumor development: an updateand outstanding questions[J]. Stem Cells, 2015, 33(8):2381-2390.
[14] Xu DD, Zhou PJ, Wang Y, et al. MiR-150 suppresses the proliferation and tumorigenicity of leukemia stem cells by targeting the nanog signaling pathway[J]. Front Pharmacol, 2016, 7(246):439.
[15] Guo D, Xu BL, Zhang XH, 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]. Oncol Rep, 2012, 28(1):241-247.
[1] 李国彬,张占成,王新颜. 上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响[J]. 山东大学耳鼻喉眼学报, 2018, 32(4): 53-57.
[2] 赵小燕,吴彩琴,任晓勇,王正辉. 白花蛇舌草多糖提取物诱导Hep-2细胞凋亡与抑制侵袭机制[J]. 山东大学耳鼻喉眼学报, 2018, 32(2): 84-87.
[3] 张彩霞,刘阳云,江文,刘庚勋,曹杭,陈琼,张纪帅. 一氧化氮对鼻咽癌CNE-2细胞株化疗增敏效应的研究[J]. 山东大学耳鼻喉眼学报, 2017, 31(5): 72-78.
[4] 李军,李晓明,吴彦桥,邸斌,苗玉花,王嘉. 累及鼻颅底区域鼻腔鼻窦骨源性良性肿瘤的鼻内镜手术治疗[J]. 山东大学耳鼻喉眼学报, 2017, 31(2): 31-36.
[5] 陈婷婷,郭其云,刘佳琪,刘丽庭. 探讨同期放化疗对儿童青少年鼻咽癌患者生活质量的影响[J]. 山东大学耳鼻喉眼学报, 2017, 31(2): 55-58.
[6] 周华群,张立庆,徐朝琪,姜盼,王愿,刘晓静,董伟达. 姜黄素联合白藜芦醇抑制人头颈部肿瘤细胞系增殖的机制研究[J]. 山东大学耳鼻喉眼学报, 2017, 31(2): 67-72.
[7] 于斌,郑青平,罗展雄,李旌,周媛媛,李中华,王磊黎,贺婵娟,石丰榕. 重组人表皮生长因子预防鼻咽癌放射性口腔黏膜炎和急性放射性皮炎的疗效观察[J]. 山东大学耳鼻喉眼学报, 2017, 31(1): 95-99.
[8] 王雅宁,耿博,李百彦,崔朝阳,王启荣. 鼻咽癌放疗后颅底骨感染坏死的治疗体会[J]. 山东大学耳鼻喉眼学报, 2016, 30(6): 42-45.
[9] 许元腾,陈瑞庆,林功标,方秀玲,俞舒娟,梁晓华,张榕. RNAi沉默PDCD4基因对人喉癌Hep-2细胞增殖及β-catenin表达的影响[J]. 山东大学耳鼻喉眼学报, 2016, 30(5): 110-114.
[10] 王君影,李晓峰. 小牛血去蛋白提取物眼用凝胶对鼻咽癌放射治疗后眼角膜内皮细胞损伤的影响[J]. 山东大学耳鼻喉眼学报, 2016, 30(1): 76-79.
[11] 赵燕恋, 卢永田, 杨继红, 张娟, 苗芳芳, 李洁萍. 鼻咽癌EGFL7的表达与肿瘤侵袭转移的关系[J]. 山东大学耳鼻喉眼学报, 2015, 29(6): 31-35.
[12] 车娟, 赵洪春, 李金玲, 王延飞. D-二聚体与鼻咽癌预后的关系[J]. 山东大学耳鼻喉眼学报, 2015, 29(5): 47-49.
[13] 陈春丽, 宋宗明, 贾新国, 周仲楼, 汪朝阳. 促红细胞生成素抑制氧化损伤诱导的人眼Müller细胞凋亡[J]. 山东大学耳鼻喉眼学报, 2015, 29(3): 65-71.
[14] 李安, 李娜, 姜彦, 于龙刚, 刘慧, 许彤, 孔祥斌. 鼻内镜手术治疗鼻-眶-颅底骨纤维异常增殖症4例[J]. 山东大学耳鼻喉眼学报, 2015, 29(2): 36-40.
[15] 刘海洋, 李甦雁, 张正培, 范巍. 改良膜分割与双手膜清除技术在23G玻璃体手术治疗增殖性糖尿病视网膜病变中的对比[J]. 山东大学耳鼻喉眼学报, 2015, 29(1): 52-55.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
网站版权所有 © 2018 《山东大学耳鼻喉眼学报》编辑部　
地址：山东省济南市山大南路27号（250100）电话：+86-0531-88366912 E-mail: ebhxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发