鼻咽癌相关内皮细胞的诱导及其鉴定  

Induction and identification of nasopharyngeal carcinoma-associated endothelial cells

作　　者：张莹 张祖荣 姚小暄 杨雨嘉 刘琦 姚茜 江川 韦正波[2] 谢莹 ZhangYing;Zhang Zurong;Yao Xiaoxuan;Yang Yujia;Liu Qi;Yao Xi;Jiang Chuan;Wei Zhengbo;Xie Ying(Institute of Life Sciences,Guangxi Medical University,Nanning 530021,China;The Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,China;Guangxi Key Laboratory of Early Prevention and Treatment of for Regional High Frequency Tumor,Ministry of Education,Nanning 530021,China)

机构地区：[1]广西医科大学生命科学研究院,南宁530021 [2]广西医科大学附属肿瘤医院,南宁530021 [3]广西区域性高发肿瘤早期防治研究教育部重点实验室,南宁530021

出　　处：《广西医科大学学报》2021年第2期229-235,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金地区基金资助项目(No.81760361);广西自然科学基金面上项目(No.2017GXNSFAA198064);区域性高发肿瘤早期防治研究教育部重点实验室自主研究项目(No.GKE-ZZ202015)。

摘　　要：目的:通过体外共培养模型,利用人鼻咽癌(NPC)细胞株(5-8F)诱导肿瘤相关人淋巴管内皮细胞(C-HLEC)及肿瘤相关人脐静脉内皮细胞(C-HUVEC)形成,分析其形态学上及相关基因表达的变化。方法:建立两种共培养体系,即嵌入式细胞共培养体系和条件培养基(CM)共培养体系,将5-8F分别与两种内皮细胞(HLEC和HUVEC)共培养,倒置相差显微镜观察共培养后内皮细胞形态学变化,实时荧光定量聚合酶链反应(qPCR)检测共培养后两种内皮细胞中肿瘤内皮标志物8(TEM8)的表达,划痕实验检测共培养后两种内皮细胞的迁移能力,小管形成实验检测共培养后两种内皮细胞成管能力。结果:与对照组比较,共培养后的两种内皮细胞,形态伸长、触角增多,变成多角形,细胞间连接疏松,细胞结构紊乱,TEM8 mRNA表达水平升高(P<0.05),且迁移能力及成管能力均明显增强(P<0.05)。结论:与NPC 5-8F细胞共培养,可促进淋巴管及血管内皮细胞向肿瘤相关内皮细胞(TECs)转化,转化后细胞迁移能力和成管能力增强、TEM8表达上调。Objective: In this study, human nasopharyngeal carcinoma cell line(5-8 F) was used to induce the formation of tumor-associated human lymphatic endothelial cells(C-HLEC) and tumor-associated human umbilical vein endothelial cells(C-HUVEC) through an in vitro co-culture model, and analyzed the changes in its morphology and related gene expression. Methods: Two co-cultivation systems, embedded cell co-cultivation system and conditioned medium co-cultivation system were established. 5-8 F was co-cultured with two endothelial cells(HLEC and HUVEC), and the morphological changes of endothelial cells after co-cultivation were observed by inverted phase contrast microscope. Real-time fluorescence quantitative polymerase chain reaction(qPCR) detected the expression of tumor endothelial markers 8(TEM8) in the two endothelial cells after co-culture, the scratch test detected the migration ability of the two endothelial cells after co-culture, and the tubule formation experiment detected the ability of the two endothelial cells to form tubes after co-culture. Results: Compared with the control group, the two kinds of endothelial cells after co-culture had elongated morphology, increased antennal,and became polygonal, loose connections between cells, disordered cell structure, increased TEM8 mRNA expression level(P<0.05). Moreover, the migration and tube formation ability were significantly enhanced(P<0.05). Conclusion: Co-cultivation with nasopharyngeal carcinoma 5-8 F cells can promote the transformation of lymphatic and vascular endothelial cells into tumor-associated endothelial cells. After transformation, the cell migration and tube formation capabilities are enhanced, and TEM8 expression is up-regulated.

关 键 词：鼻咽癌 肿瘤相关内皮细胞 迁移 肿瘤内皮标志物8(TEM8) 血管生成 

分 类 号：R739.63[医药卫生—肿瘤]