Biphasic effect of thrombospondin-1 (TSP-1) in the regulation of angiogenesis in human breast carcinoma
Biphasic effect of thrombospondin-1 (TSP-1) in the regulation of angiogenesis in human breast carcinoma
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DOI：
10.1042/bst024368s
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发表时间：
1996-08-01
影响因子：
3.9
通讯作者：
Schor, AM
中科院分区：
文献类型：
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作者：
Pazouki, S;Pendleton, N;Schor, AM
TSP-1是一种与血管生成有关的大细胞外基质糖蛋白。其具体作用尚不清楚，因为刺激和抑制作用已在动物模型中得到证实（1,2）。血管生成不能在人类肿瘤中直接测量，然而，组织切片中微血管的密度通常被用作这一过程的指标。尽管血管生成对肿瘤生长和传播很重要，但微血管密度作为乳腺癌预后指标的价值仍存在争议（3-5）。为了评估tsp - 1在血管生成中的可能作用，我们在切除的人乳腺癌中量化了血管状况和tsp - 1的表达。采用四种不同的方法对组织切片进行血管性血友病因子（vWF）抗体免疫染色。这些方法分别是：(a)平均微血管密度（a- mvd），(b)最高微血管密度（h-MVD），(c)微血管体积（MW）和(d)染色区域图像分析（vWF area）。a- mvd和h-MVD通过计算覆盖0.476 mm2面积的网格内的血管来评估。为了测量a-MVD，在15-20个随机区域内对血管进行计数。为了测量h-MVD，对肿瘤切片进行扫描，寻找血管密度最高的区域；然后统计该区域5个油田的船舶数量，取最高值。在这两种情况下，计数都以每平方毫米的血管数表示。MVV的测量采用目镜光栅点计数法，目镜光栅包含100个点；15个随机字段（1500点）在每个部分被计算。通过(a)使用32p标记的cDNA探针对人内皮tsp - 1进行northern blotting （nb-TSP）和(b)使用地高辛标记的RNA探针进行原位杂交（ish-TSP）来测定tsp - 1 mRNA的表达。nb-TSP采用密度扫描法定量，ish-TSP采用形态计量法对15个随机场进行点计数。
TSP-1 is a large extracellular matrix glycoprotein implicated in angiogenesis. Its specific role is not clear, as both stimulatory and inhibitory effects have been demonstrated in animal models (I, 2). Angiogenesis cannot be measured directly in human tumours, however, the density of the microvasculature in tissue sections has been commonly used as an index of this process. Although angiogenesis is important for tumour growth and dissemination, the value of microvascular density as a prognostic indicator in breast carcinoma remains controversial (3-5).In order to assess the possible role of TSP-I in angiogenesis, we have quantitated vascularity and TSP-I expression in resected human breast carcinomas. Vascularity was assessed by four different methods following immunostaining of histological sections with antibodies to Von Willebrand factor (vWF). These methods were:(a) average microvascular density (a-MVD),(b) highest microvascular density (h-MVD),(c) microvascular volume (MW) and (d) image analysis of stained area (vWF Area). a-MVD and h-MVD were assessed by counting the vessels that were contained within a grid covering an area of 0.476 mm2. To measure a-MVD, vessels in 15-20 random fields across the section were counted. To measure h-MVD, the tumour sections were scanned for the area of highest vascular density; the number of vessels in 5 fields in this area were then counted and the highest value was taken. In both cases the counts were expressed as number of vessels per square millimetre. MVV was measured by point counting using an eyepiece graticule which contained 100 points; 15 random fields (1500 points) were counted across each section. TSP-I mRNA expression was determined by (a) northern blotting (nb-TSP) using a 32P-labelled cDNA probe to human endothelial TSP-I, and (b) in situ hybridisation (ish-TSP) using a digoxigenin-labelled RNA probe. nb-TSP was quantitated by densitometric scanning and ish-TSP was quantitated morphometrically by point counting of 15 random fields.