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An immuno-scanning electron microscope study of the tumor-stromal border in the invasive edge of colorectal cancers

结直肠癌浸润边缘肿瘤间质边界的免疫扫描电子显微镜研究

基本信息

批准号：
06671309
负责人：
MIURA Seiji
金额：
$ 1.41万
依托单位：
Teikyo University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

To elucidate three-dimensional relationships among the tumor gland, basement membrane, and fibrous stroma in the invasive edge of colorectal cancers, we have attempted several techniques to identify types of collagen fibers by using immuno-scanning electron microscopy (immune-SEM) . During the term of project, immuno-gold labeling of the specimens and observation by immune-SEM with both backscattered and secondary electron imaging, and Texas-red labeling of the similarly prepared specimens and observation by confocal laser-scanning microscopy (CLSM) with three-dimensional image reconstruction were performed, where CLSM being performed to estimate the gold-labeled site on immune-SEM.Stromel collagen fibers, identified as type I,tended to form thick bundles and run parallel to the acinar outer surface. Few fibrous connections were noted between the basement membrane of the cancer acinus (or acinus without basement membranes) and the stromal collagen fibers. Type IV collagen fibers, forming the basement membrane of cancer acini, were observed as waving and densely-arranged bundles of fibers but not wide-meshed, being occasionally discontinuous or absent. In contrast, the basement membrane of the normal crypt formed a wide-meshed and continuous lattice of fibers, and numerous fibrous connections with the interstitial cellagen fibers were noted.Conclusions : 1. The basement membrane of the tumor acini may neither provide a firm anchorage to cancer cells nor be suitable for the penetration of substances, in contrast to that of the normal crypt. The theory that it is a newly-deposited structure, rather than the preexisting basement membrane destroyed by tumor cells, might be supported by the present results. 2. Application of immune-SEM in tissue (not cytologic) level may provide a new way for studying the tumor-stromal interface, which, recently, has been studied by so many investigators from different fields of onocology.

为了阐明结直肠癌侵袭边缘的肿瘤腺体、基底膜和纤维间质之间的三维关系，我们尝试了几种利用免疫扫描电子显微镜（immune-SEM）识别胶原纤维类型的技术。在项目期间，对样品进行免疫金标标记和免疫扫描电镜观察，同时进行背散射和二次电子成像，并对类似制备的样品进行德克萨斯红标记和共聚焦激光扫描显微镜（CLSM）观察，并进行三维图像重建，其中使用CLSM来估计免疫扫描电镜上的金标位点。Stromel胶原纤维，鉴定为I型，倾向于形成粗束，平行于腺泡外表面。癌腺泡基底膜（或无基底膜的腺泡）与间质胶原纤维之间很少有纤维连接。形成癌腺基底膜的IV型胶原纤维呈波状、密集排列的纤维束，但不宽网，偶有不连续或缺失。相比之下，正常隐窝的基底膜形成了一个宽网格的连续纤维晶格，并与间质纤维素纤维有许多纤维连接。结论：1。与正常隐窝相比，肿瘤腺泡的基底膜既不能为癌细胞提供牢固的锚定，也不适合物质的渗透。这是一种新沉积的结构，而不是先前存在的基底膜被肿瘤细胞破坏的理论，可能得到本研究结果的支持。2. 免疫扫描电镜在组织（非细胞学）水平上的应用，为肿瘤-基质界面的研究提供了一种新的途径，近年来已受到肿瘤学不同领域研究者的广泛研究。