MiR-150调控Nanog对鼻咽癌侧群细胞增殖、侵袭的影响  

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

作　　者：周毅波[1] 龚小蓉 于锋[1] ZHOU Yibo GONG Xiaorong YU Feng(Guangzhou 12th People＇s Hospital, Guangzhou Hospital of Otolaryn- gology Head and Neck Surgery, Institute of Otolaryngology Head and Neck Surgery of Guangzhou Medical University, Guangzhou 510620, Guangdong, China)

机构地区：[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

出　　处：《山东大学耳鼻喉眼学报》2017年第5期79-84,共6页Journal of Otolaryngology and Ophthalmology of Shandong University

基　　金：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)

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,w estern blotting检测Nanog的表达情况;进一步通过CCK-8实验、Transw all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,M iR-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M iR-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、Transw ell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M iR-150与Nanog呈高表达,M iR-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。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.

关 键 词：鼻咽癌 MiR-150 SP细胞 NSP细胞 增殖 侵袭 

分 类 号：R739.63[医药卫生—肿瘤]