口腔癌患者循环外泌体促进舌癌细胞CAL27增殖、迁移、侵袭中的作用
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摘要
目的:探讨口腔癌患者外周血中循环外泌体(CEXO)对口腔癌细胞增殖、侵袭中作用。方法:纳入口腔鳞状细胞癌患者20例。梯度离心法收集口腔癌患者外周血中循环外泌体,以透射电镜和蛋白印迹法鉴定收集所得的沉淀。将收集的外泌体加到舌癌细胞CAL27的培养中,MTT法检测细胞增殖能力,划痕愈合试验检测细胞迁移能力,Transwell小室法检测细胞侵袭能力,最后利用CAL27细胞裸鼠移植瘤模型,检测口腔癌患者循环外泌体在体内对癌细胞增殖能力的影响。结果:在体外实验中,口腔癌患者循环外泌体在体外可促进舌癌细胞增殖、迁移、侵袭;在裸鼠移植瘤模型中亦可促进口腔癌细胞增殖。结论:口腔癌患者外周血循环外泌体可促进口腔癌细胞的增殖、迁移和侵袭。
Objective: To investigate the effects of circulating exosomes derived from oral cancer patients on the proliferation,migration and invasion of tongue cancer CAL27 cells. Methods: Circulating exosomes were isolated from the peripheral blood of 20 oral cancer patients using gradient centrifugation. The characteristics of the exosome was studied by transmission electron microscope( TEM) and Western blot. After treatment with circulating exosomes,CAL27 cell proliferation was evaluated with MTT assay,migration was observed by cell wound scratch assay and the invasion of CAL27 cells was tested by Transwell assay. CAL27 cell bearing nude mice were used to estimate the effect of circulating exosome on CAL27 cell proliferation and metastasis in vivo. Results: The proliferation,migration and invasion of CAL27 cells were enhanced by circulating exosomes from oral cancer patients. In vivo study showed that the tumorigenesis and metastasis of CAL27 cells were enhanced by circulating exosomes. Conclusion: Circulating exosomes from oral cancer patients can promote the proliferation,migration,invasion and metastasis of CAL27 cells.
Objective: To investigate the effects of circulating exosomes derived from oral cancer patients on the proliferation,migration and invasion of tongue cancer CAL27 cells. Methods: Circulating exosomes were isolated from the peripheral blood of 20 oral cancer patients using gradient centrifugation. The characteristics of the exosome was studied by transmission electron microscope( TEM) and Western blot. After treatment with circulating exosomes,CAL27 cell proliferation was evaluated with MTT assay,migration was observed by cell wound scratch assay and the invasion of CAL27 cells was tested by Transwell assay. CAL27 cell bearing nude mice were used to estimate the effect of circulating exosome on CAL27 cell proliferation and metastasis in vivo. Results: The proliferation,migration and invasion of CAL27 cells were enhanced by circulating exosomes from oral cancer patients. In vivo study showed that the tumorigenesis and metastasis of CAL27 cells were enhanced by circulating exosomes. Conclusion: Circulating exosomes from oral cancer patients can promote the proliferation,migration,invasion and metastasis of CAL27 cells.
引文
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[5]Languino LR,Singh A,Prisco M,et al.Exosome-mediated transfer from the tumor microenvironment increases TGFβsignaling in squamous cell carcinoma[J].Am J Transl Res,2016,8(5):2432-2437.
[6]Ratajczak J,Wysoczynski M,Hayek F,et al.Membranederived microvesicles:Important and underappreciated mediators of cell-to-cell communication[J].Leukemia,2006,20(9):1487-1495.
[7]Ridder K,Keller S,Dams M,et al.Extracellular vesiclemediated transfer of genetic information between the hematopoietic system and the brain in response to inflammation[J].PLo S Biol,2014,12(6):e1001874.
[8]Min L,Shen J,Tu C,et al.The roles and implications of exosomes in sarcoma[J].Cancer Metastasis Rev,2016,35(3):377-390.
[9]Zhang L,Zhang S,Yao J,et al.Microenvironment-induced PTEN loss by exosomal microRNA primes brain metastasis outgrowth[J].Nature,2015,527(7576):100-104.
[10]Hoshino A,Costa-Silva B,Shen TL,et al.Tumour exosome integrins determine organotropic metastasis[J].Nature,2015,527(7578):329-335.
[11]Principe S,Hui AB,Bruce J,et al.Tumor-derived exosomes and microvesicles in head and neck cancer:Implications for tumor biology and biomarker discovery[J].Proteomics,2013,13(10-11):1608-1623.
[12]Vader P,Mol EA,Pasterkamp G,et al.Extracellular vesicles for drug delivery[J].Adv Drug Deliv Rev,2016,106(Pt A):148-156.
[13]Rider MA,Hurwitz SN,Meckes DG Jr.Extra PEG:A polyethylene glycol-based method for enrichment of extracellular vesicles[J].Sci Rep,2016,6:23978.
[14]杨子楠,魏继武.外泌体在肿瘤发展中的研究进展[J].肿瘤,2011,31(6):565-569.
[15]Yang N,Li S,Li G,et al.The role of extracellular vesicles in mediating progression,metastasis and potential treatment of hepatocellular carcinoma[J].Oncotarget,2017,8(2):3683-3695.
[16]Matsumoto Y,Kano M,Akutsu Y,et al.Quantification of plasma exosome is a potential prognostic marker for esophageal squamous cell carcinoma[J].Oncol Rep,2016,36(5):2535-2543.
[17]刘善廷,吴俊福,郑荣寿,等.2009年中国口腔癌和咽喉癌发病与死亡情况分析[J].中华预防医学杂志,2013,47(7):586-591.
[18]邱蔚六.邱蔚六口腔颌面外科学[M].上海:上海科学技术出版社,2008:389.
[19]韩晟,陈衍,谢诚,等.576例口腔癌患者的流行病危险因素研究[J].实用口腔医学杂志,2012,28(1):74-78.
[20]金龙,杨宏宇,杨辉俊,等.NOD2上调自噬抑制舌鳞癌细胞增殖和迁移[J].实用口腔医学杂志,2015,31(3):352-356.
[21]闫晋,谢利明,沈国芳,等.GSTM1基因、吸烟及饮酒与亚洲人群口腔癌易感性的Meta分析[J].实用口腔医学杂志,2014,30(4):515-521.