Sinha A A, Gleason D F, Staley N A, Wilson M J, Sameni M, Sloane B F
Research Service, Veterans Affairs Medical Center, Minneapolis, Minnesota, USA.
Anat Rec. 1995 Mar;241(3):353-62. doi: 10.1002/ar.1092410309.
Angiogenesis (or neovascularization) is required for the growth of solid organ tumors and precedes invasion of the adjacent stroma by neoplastic cells. We investigated the relative density and distribution of cathepsin B (CB) immunostained microvessels (i.e., small blood vessels and capillaries) in benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN), and prostatic adenocarcinoma (CAP) by immunocytochemical localization of an antibody directed against a cathepsin B-derived synthetic peptide (Syn-CB).
We studied 16 formalin-fixed, prostatectomy specimens that were embedded in paraffin/paraplast for histological examination by hematoxylin and eosin and immuno-localization of the Syn-CB antibody. Selected paraformaldehyde-fixed specimens were embedded in K4M Lowicryl or LRWhite resins. We localized the antibody in thin sections using immunoelectron microscopy techniques.
Eight patients had BPH [4 patients with BPH alone, 2 with BPH and PIN, and 2 with BPH and CAP]. Ten cancer cases included one with Gleason histologic score 4, two with score 6, four with score 7, and three with score 8. In CAP cases, Gleason score 6 and 7 tumors had more microvessels than the score 4 or 8 tumors. In both BPH and CAP cases, the antibody was localized chiefly in the endothelial cells of microvessels, but occasionally in ductal and glandular epithelial cells. Ultrastructurally, CB-immunoreactive gold particles were markedly increased at the luminal and basal plasma membrane surfaces and folds of endothelial cells in neoplastic prostate, but not in the endothelial cells of BPH. Furthermore, the presence of CB localizing gold particles in collagen and smooth muscle fibers near the microvessels indicated leakage of the enzyme in prostatic stroma of neoplastic prostate. Similar leakage was not observed in BPH. Morphometric analysis showed that the relative density of microvessels increased two to three times in cancer patients when compared to patients with BPH alone. Our study also indicated that BPH associated with PIN or CAP had an increased density of microvessels when compared to BPH alone.
Our study showed that the relative density and distribution of microvessels are the most important features of neovascularization in prostatic tumors. The relative density of microvessels increased in PIN and CAP when compared to BPH alone. Although the localization of CB is associated with lysosomes of endothelial cells in both BPH and CAP, there is a greater association of CB with the plasma membranes of endothelial cells in CAP than BPH. Immunoelectron microscopy provided evidence that CB might be involved in dissolution of basement membranes in neoplastic tumors during angiogenesis. CB localization has the potential of defining a role for this protease in degradation of extracellular matrix constituents during early steps of angiogenesis.
实体器官肿瘤的生长需要血管生成（或新生血管形成），且肿瘤细胞侵袭相邻基质之前就已发生血管生成。我们通过针对组织蛋白酶B衍生合成肽（Syn-CB）的抗体进行免疫细胞化学定位，研究了良性前列腺增生（BPH）、前列腺上皮内瘤变（PIN）和前列腺腺癌（CAP）中组织蛋白酶B（CB）免疫染色微血管（即小血管和毛细血管）的相对密度和分布。
我们研究了16例福尔马林固定的前列腺切除标本，这些标本用石蜡/石蜡包埋，用于苏木精和伊红组织学检查以及Syn-CB抗体的免疫定位。选择的多聚甲醛固定标本用K4M Lowicryl或LRWhite树脂包埋。我们使用免疫电子显微镜技术在薄切片中定位抗体。
8例患者患有BPH[4例单纯BPH，2例BPH合并PIN，2例BPH合并CAP]。10例癌症病例包括1例Gleason组织学评分为4分，2例评分为6分，4例评分为7分，3例评分为8分。在CAP病例中，Gleason评分为6分和7分的肿瘤比评分为4分或8分的肿瘤微血管更多。在BPH和CAP病例中，抗体主要定位于微血管的内皮细胞，但偶尔也定位于导管和腺上皮细胞。超微结构上，CB免疫反应性金颗粒在肿瘤性前列腺内皮细胞的腔面和基底质膜表面及褶皱处明显增加，而在BPH的内皮细胞中未增加。此外，微血管附近的胶原和平滑肌纤维中存在CB定位金颗粒表明该酶在肿瘤性前列腺的前列腺基质中渗漏。在BPH中未观察到类似的渗漏。形态计量分析表明，与单纯BPH患者相比，癌症患者微血管的相对密度增加了两到三倍。我们的研究还表明，与单纯BPH相比，合并PIN或CAP的BPH微血管密度增加。
我们的研究表明，微血管的相对密度和分布是前列腺肿瘤新生血管形成的最重要特征。与单纯BPH相比，PIN和CAP中微血管的相对密度增加。虽然CB的定位在BPH和CAP中均与内皮细胞的溶酶体相关，但与BPH相比，CB在CAP中与内皮细胞质膜的相关性更大。免疫电子显微镜提供了证据，表明CB可能在血管生成过程中参与肿瘤性肿瘤基底膜的溶解。CB定位有可能确定这种蛋白酶在血管生成早期细胞外基质成分降解中的作用。
