人脉络膜黑色素瘤细胞株OCM-1的三维培养及血管生成拟态机制的研究

发布时间：2019-02-12 13:21

【摘要】：目的通过体外三维培养人脉络膜黑色素瘤细胞株OCM-1,观察其生成血管生成拟态(vasculogenic mimicry,VM)时,PI3K、Eph A2蛋白的表达,探讨VM发生的可能机制。方法体外三维培养OCM-1细胞,于三维培养7 d进行过碘酸-雪夫反应(periodic acid schiff reaction,PAS)染色,观察PAS阳性的环状结构的形成(即其形成VM的能力),于1 d、4 d、7 d进行PI3 K和Eph A2免疫组织化学染色并观察其结果。结果OCM-1细胞三维培养4 d,细胞大部分呈多角形,胞浆较丰富,核圆形,可见核仁,少部分肿瘤细胞长梭形,可见3~5个长梭形细胞彼此连接围成半环状结构。7 d后,肿瘤细胞几乎均变成长梭形,沿着胶原支架生长,细胞伸出长的突起,彼此之间相互接触围成环状结构。PAS染色可见瘤细胞大多呈弥散排列,肿瘤细胞模仿机体血管生成而形成瘤细胞条索并围成管道,一层细胞外基质(PAS阳性物质)围成环状结构。与培养1 d的细胞相比,培养4 d和7 d的PI3K和Eph A2蛋白表达量均明显增加(均为P0.05),并随着时间的推移增加(7 d4 d1 d,P0.05)。结论Eph A2和PI3 K可能在三维培养人脉络膜黑色素瘤细胞株OCM-1的VM形成过程中具有重要作用。
[Abstract]:Objective to investigate the expression of PI3K,Eph A2 protein during angiogenic mimicry (vasculogenic mimicry,VM) of human choroidal melanoma cell line OCM-1, in vitro, and to explore the possible mechanism of VM. Methods OCM-1 cells were cultured in vitro for 7 days. The formation of PAS positive ring structure (that is, its ability to form VM) was observed by (periodic acid schiff reaction,PAS staining. PI3 K and Eph A2 immunohistochemical staining were performed on 7 days and the results were observed. Results for 4 d, most of OCM-1 cells were polygonal, with abundant cytoplasm, round nucleus, long fusiform of a few tumor cells, and 3 ~ 5 long fusiform cells connected with each other to form a semicircular structure 7 days later. Almost all tumor cells become fusiform, grow along collagen scaffolds, the cells extend long protuberances, and contact with each other to form a circular structure. PAS staining shows that most of the tumor cells are distributed. Tumor cells mimic the angiogenesis of the body and form tumor cell lines and enclosing pipes, and a layer of extracellular matrix (PAS positive substances) is surrounded by a circular structure. Compared with the cells cultured for 1 day, the expression of PI3K and Eph A2 protein increased significantly at 4 and 7 days (P0.05), and increased with the passage of time (P0.05 on 7 d 4 d 1 d). Conclusion Eph A2 and PI3 K may play an important role in the formation of VM in human choroidal melanoma cell line OCM-1.
【作者单位】：天津中医药大学第一附属医院眼科;天津市眼科医院;天津医科大学眼科医院;
【分类号】：R739.7

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