外泌体介导的miR-577在幼儿复发性喉乳头状瘤中的表达及其对细胞增殖和侵袭的影响

Expression of Exosome Mediated miR-577 in Children with Recurrent Laryngeal Papilloma and Its Effect on Proliferation and Invision of Laryngeal Papilloma Cells

胡彬雅;李赟;敬云龙;赵斯君;

1:湖南省儿童医院耳鼻咽喉头颈外科

摘要
目的探讨miR-577在幼儿复发性喉乳头状瘤中的表达及其对细胞增殖、侵袭的影响和潜在分子机制。方法收集2016年6月至2019年12月确诊的30例幼儿喉乳头状瘤组织样本及其相匹配且远离病灶经病理检查证实的正常喉组织样本,并选取人正常喉上皮细胞株和喉乳头状瘤细胞(分为miR-577过表达组、阴性对照组及空白对照组)进行研究。采用实时荧光定量PCR(qRT-PCR)分别检测30例喉乳头状瘤组织、配对正常喉组织、正常喉上皮细胞和喉乳头状瘤细胞中miR-577相对表达;通过Lipofectamine(TM)3000脂质体转染试剂盒将miR-577 mimic模拟物转染至喉乳头状瘤细胞构建miR-577过表达细胞系,采用CCK-8法、Tanswell实验分别检测miR-577对喉乳头状瘤细胞增殖、侵袭的影响;采用Western blot检测miR-577对喉乳头状瘤细胞侵袭相关蛋白(MMP-1、MMP-9)表达及Notch通路相关蛋白(Notch、DSL)表达的影响。结果幼儿喉乳头状瘤组织中miR-577相对表达量(0.134±0.026)显著低于正常喉组织(0.823±0.056)(P<0.001);喉乳头状瘤细胞中miR-577相对表达量(0.256±0.026)显著低于正常喉上皮细胞(0.962±0.072)(P<0.001);与阴性对照组相比,转染miR-577 mimic后喉乳头状瘤细胞增殖能力(1.248±0.103 vs 0.902±0.094)和细胞侵袭率(71.016±5.869 vs 42.867±4.795)明显降低(P<0.05);侵袭相关蛋白MMP-1(1.011±0.124 vs 0.417±0.056)、MMP-9(1.012±0.155 vs 0.502±0.058)表达抑制(P<0.01)。喉乳头状瘤细胞中Notch、DSL表达显著上调(P<0.05);转染miR-577 mimic后喉乳头状瘤细胞中Notch通路相关蛋白Notch、DSL表达明显抑制(P<0.05)。共转染miR-577 mimic和pcDNA3.1-Notch后喉乳头状瘤细胞增殖能力和细胞侵袭率较单纯转染miR-577 mimic组明显增加(P<0.05);共转染pcDNA3.1-Notch逆转了miR-577 mimic对喉乳头状瘤细胞增殖、侵袭能力的抑制作用。结论幼儿喉乳头状瘤组织及喉乳头状瘤细胞中miR-577显著低表达,过表达miR-577显著抑制喉乳头状瘤细胞的增殖和侵袭;Notch是miR-577的潜在靶点,miR-577可能通过与Notch的3'UTR区结合进而调控Notch信号通路来抑制喉乳头状瘤细胞的增殖和侵袭。
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关键词
miR-577;喉乳头状瘤;增殖;侵袭;Notch信号通路
基金项目(Foundation):
湖南省卫生计生委科研基金课题(B2011-093)
作者
胡彬雅;李赟;敬云龙;赵斯君;
参考文献

1 Omura K,Nomura K,Aoki S,et al.Resection of inverted papilloma in nasal cavity with transseptal access and crossing multiple incisions minimizes bleeding and reveals the tumor pedicle[J].Auris,Nasus,Larynx,2019,245(11):412-419.

2 胡彬雅,赵斯君,任基浩,等.儿童喉乳头状瘤组织中Toll样受体4及激活蛋白-1表达及相关性研究[J].现代肿瘤医学,2018,245(11):42-45.

3 Denzer K,Kleijmeer MJ,Heijnen HF,et al.Exosome:from internal vesicle of the multivesicular body to intercellular signaling device[J].J Cell Sci,2000,19(7):3365-3374.

4 Saadatpour L,Fadaee E,Fadaei S,et al.Glioblastoma:exosome and microRNA as novel diagnosis biomarkers [J].Cancer Gene Ther,2016,23(12):415-418.

5 Liu D,Zhang C,Li X,et al.MicroRNA-567 inhibits cell proliferation,migration and invasion by targeting FGF5 in osteosarcoma[J].Excli Journal,2018,17(11):102-112.

6 Wang Y,Lu Z,Wang N,et al.Long noncoding RNA DANCR promotes colorectal cancer proliferation and metastasis via miR-577 sponging[J].Exp Mol Med,2018,50(5):57-65.

7 华涛,许家玲.miR-577通过介导Notch信号通路抑制肺癌细胞的增殖、侵袭和耐药性[J].临床肺科杂志,2020,25(2):177-182.

8 郭梦,石瑞,毛维.miR-577对胶质瘤细胞上皮-间质转化的调控作用及机制[J].中华实用诊断与治疗杂志,2019,33(12):1158-1161.

9 Quezada C,Torres A,Niechi I,et al.Role of extracellular vesicles in glioma progression[J].Mol Aspects Med,2018,60(8):38-51.

10 Becker A,Thakur BK,Weiss JM,et al.Extracellular vesicles in cancer:cell-to-cell mediators of metastasis[J].Cancer Cell,2016,30(6):836-848.

11 Zhu L,Oh JM,Gangadaran P,et al.Targeting and therapy of glioblastoma in a mouse model using exosomes derived from natural killer cells[J].Front Immunol,2018,93(9):1-13.

12 Bigas A,Espinosa L.Notch signaling in cell-cell communication Pathways[J].Current Stem Cell Reports,2016,2(4):349-355.

13 Ming L,Qian Z,Liang W,et al.Serum miR-499 as a novel diagnostic and prognostic biomarker in non-small cell lung cancer[J].Oncology Reports,2014,31(4):1961-1967.

14 Zhang W,Shen C,Li C,et al.miR-577 inhibits glioblastoma tumor growth via the Wnt signaling pathway[J].Molecular Carcinogenesis,2016,55(5):575-585.

15 Wang LY,Li B,Jiang HH,et al.Inhibition effect of miR-577 on hepatocellular carcinoma cell growth via targeting β-catenin[J].Asian Pac J Trop Med,2015,8(11):923-929.

16 Yin C,Mou Q,Pan X,et al.MiR-577 suppresses epithelial-mesenchymal transition and metastasis of breast cancer by targeting Rab25[J].Thoracic Cancer,2018,9(4):472-479.

17 Zhou F,Pan Z,Shen F,et al.Long noncoding RNA DLX6-AS1 functions as a competing endogenous RNA for miR-577 to promote malignant development of colorectal cancer[J].Eur Rev Med Pharmacol Sci,2019,23(9):3742-3748.

18 Majidinia M,Darband SG,Kaviani M,et al.Cross-regulation between Notch signaling pathway and miRNA machinery in cancer[J].Dna Repair,2018,66(11):30-41.

19 Aster JC,Pear WS,Blacklow SC.The varied roles of Notch in cancer[J].Annu Rev Pathol,2017,12(1):245-255.

20 Mamaeva V,Niemi R,Beck M,et al.Inhibiting notch activity in breast cancer stem cells by glucose functionalized nanoparticles carrying γ-secretase inhibitors[J].Molecular Therapy,2016,24(5):926-936.

21 Kim B,Stephen SL,Hanby AM,et al.Chemotheraphy induce Notch1 dependent MRP1 up-regulation,inhibition of which sensitizes breast cancer cells to chemotheraphy[J].BMC Cancer,2015,15(1):1-12.

22 袁勇,胡杨,赵雍凡,等.Notch信号通路相关蛋白在食管鳞状细胞癌中的表达[J].中华胃肠外科杂志,2015,18(9):909-913.

23 Yen WC,Fischer MM,Axelrod F,et al.Targeting Notch signaliing with a Notch2/Notch3 antagonist (Tarextumab) inhibits tumor growth and decreases tumor-initiating cell frequency [J].Clin Cancer Res,2015,21(9):2084-2095.

24 Yong F,Cao T,Guo WC,et al.Cinobufagin induces apoptosis of osteosarcoma cells through inactivation of Notch signaling[J].Eur J Pharmacol,2017,79(4):77-84.

25 Yuan X,Wu H,Xu H,et al.Notch signaling:an emerging therapeutic target for cancer treatment[J].Cancer Letters,2015,369(1):20-27.

26 Lim JS,Ibaseta A,Fischer MM,et al.Intratumoural heterogeneity generated by Notch signalling promotes small-cell lung cancer[J].Nature,2017,545(7):360-341.

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