MiR-150调控Nanog对鼻咽癌侧群细胞增殖、侵袭的影响

doi:10.6040/j.issn.1673-3770.0.2017.321

摘要/Abstract

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

周毅波,龚小蓉,于锋. MiR-150调控Nanog对鼻咽癌侧群细胞增殖、侵袭的影响[J]. 山东大学耳鼻喉眼学报, 2017, 31(5): 79-84.

ZHOU Yibo, GONG Xiaorong, YU Feng. Effects of miR-150 targeted to Nanog on proliferation and invasion of nasopharyngeal carcinoma side population cells.[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2017, 31(5): 79-84.

参考文献

[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.

相关文章 15

[1]	林小雪,林葆睿,李佩珊,卢标清. 电子鼻咽镜联合窄带成像技术在鼻咽癌中医辨证中的应用[J]. 山东大学耳鼻喉眼学报, 2026, 40(3): 40-46.
[2]	王盛,李银丹,杨金姬. microRNA在鼻咽癌中的作用及其研究进展[J]. 山东大学耳鼻喉眼学报, 2025, 39(5): 125-131.
[3]	邱前辉,肖旭平,杨钦泰,叶菁,邓泽义,王德生,谭国林,蒋卫红,卢永田,唐隽,石照辉,邓晓聪,刘遗斌,王跃武,段传志,杜德坤,白小欣,陈文伙,莫立根,蔡楚伟,曾鹏,何旭英,杨一梅,赵洲洋,陈健龙,赵充,林志雄,李先明,李曙平,陈冬平,陈勇,黄莹,陈春燕,韩非,黄理金,瞿申红. 鼻咽癌治疗后并发颈动脉爆裂综合征的临床处理专家共识[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 1-18.
[4]	王思权,朱洪申,张晓斌,赵洲洋,马跃,杨一梅,黄理金. 放射治疗后鼻咽癌患者单侧颈内动脉栓塞术后脑卒中及颅神经麻痹的相关因素分析[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 19-25.
[5]	黄巧,任毅,侯涛,廖行伟,朱子昂,詹晓琳,刘盈,尹时华. 鼻咽癌组织中EphB2表达及与临床病理特征的相关性[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 26-30.
[6]	孙春晓,王文晴,岳田,刘济生. 高低累积顺铂剂量同步放化疗治疗鼻咽癌的疗效分析[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 31-41.
[7]	徐飞,朱光熹,王可心. 基于决策树算法构建鼻咽癌患者放疗后发生放射性口腔黏膜炎风险的预测模型[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 42-48.
[8]	王再兴,唐志元,李定波,石照辉,曾宪海,张秋航. 鼻咽癌放疗后肿瘤复发及颅底骨坏死引起颈内动脉破裂的治疗方案[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 49-58.
[9]	孙芳,谢楚波,邱前辉. 营养指标对鼻咽癌放射性颅底坏死患者创面修复影响的回顾性分析[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 59-68.
[10]	朱瑞楷,吴家荣,孙芳,谢楚波,邱前辉. 基于计算机断层扫描血管造影术评估鼻咽癌放疗后引起颈内动脉狭窄状况及其影响因素的研究[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 77-84.
[11]	覃德波,薛建成,杨文月,胡兵,陈涛,俞艳萍,孟庆国,孙焕吉,苗北平,卢永田. 鼻咽癌诊疗变革:生物标志物与鼻内镜手术协同推进早期治疗发展[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 85-92.
[12]	吴家荣,邱前辉. 颅底筋膜组织在早期复发性鼻咽癌内镜手术中的临床意义[J]. 山东大学耳鼻喉眼学报, 2025, 39(4): 108-113.
[13]	杨鸣,刘雪霞,张华. m6A识别蛋白IGF2BPs家族在头颈肿瘤中的研究进展[J]. 山东大学耳鼻喉眼学报, 2025, 39(3): 153-161.
[14]	吴敏,李正阳,孟杰,叶惠平. 程序性细胞死亡的分子机制和其在鼻咽癌中的作用[J]. 山东大学耳鼻喉眼学报, 2025, 39(2): 152-157.
[15]	张茂华,魏日富,朱忠寿,刘平,高尚,李慧凤. LncRNA PCAT-1对鼻咽癌细胞生物学行为及化疗敏感性的影响[J]. 山东大学耳鼻喉眼学报, 2025, 39(1): 68-76.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed