Three-dimensional quantification of tumor microenvironment in tongue squamous cell carcinoma

本研究旨在获得舌鳞状细胞癌（SCC）微环境中癌细胞浸润模式、血管网络和成纤维细胞/巨噬细胞聚集的三维定量信息。在批准的2年期间，使用从30例SCC制备的100个连续组织切片实现3D重建。采用免疫组化结合抗体（细胞角蛋白、Ki-67、CD 31、CD 105、CD 34、CD 68、αSMA、S100 A4）对癌实质和间质的表型和结构进行表征。所得到的3D构建体提供了足够的空间分辨率以抓住单个细胞、肿瘤血管的异常发散和迂曲、癌血管侵袭和基质细胞。我们的经验证明，三维重建提供了一个重要的工具，以验证癌细胞，血管内皮细胞，肌成纤维细胞（CAF）和巨噬细胞（TAM）的空间关联。

The present study aimed at gaining 3D quantitative information about carcinoma invasion pattern, vascular network and accumulating fibroblasts/macrophages in the microenvironment of tongue squamous cell carcinoma(SCC). During the approved 2-year period, 3D reconstruction was achieved using 100 serial histological sections that were prepared from 30 cases of SCC. Phenotypes and architecture of carcinoma parenchyma and stroma were characterized by stepwise immunohistochemistry in combination with antibodies(cytokeratin, Ki-67, CD31, CD105, CD34, CD68,αSMA, S100A4). The resulting 3D constructs provided spatial resolution enough to grasp individual cells, aberrant divergence and tortuosity of tumor vessels, carcinoma vascular invasion and stromal cells. Our experience proved that 3D reconstruction offers an essential tool to validate the spatial association of cancer cells, vascular endothelial cells, myofibroblasts(CAF) and macrophages(TAM).