Immunohistochemistry in the evaluation of neovascularization in tumor xenografts
Immunohistochemistry in the evaluation of neovascularization in tumor xenografts
复制标题
DOI：
10.1080/10520290802451085
复制
发表时间：
2008-01-01
影响因子：
1.6
通讯作者：
Siegal, G. P.
中科院分区：
文献类型：
--
作者：
Wang, D.;Stockard, C. R.;Siegal, G. P.
众所周知，血管生成或新生血管在导致转移的肿瘤进展中起着重要作用。CD31或因子VIII相关抗原(F VIII RAg)免疫组织化学被广泛用于定量移植转化的人类细胞系的免疫低下动物模型中的肿瘤新生血管的实验研究。然而，量化可能会受到用于测量血管形成的方法学的变化的影响，包括抗体选择、抗原修复(AR)预处理和评估技术。为了进一步研究这一点，我们用抗CD31和F VIII RAG免疫组织化学染色研究了五种不同的人肿瘤移植瘤和一种小鼠同基因肿瘤的微血管密度(MVD)和微血管染色强度。对不同的AR方法也进行了评价。用0.5M Tris(PH 10)缓冲液最大回收CD31，用0.05%胃酶处理组织切片可最大回收F RAG。对于每个优化的检索条件，抗CD31抗体突出小血管优于F VIII RAG。此外，CD31微血管密度明显大于F VIII RAG装饰血管(p<；0.001)。在研究血管生成时，抗体和AR方法的选择对免疫组织化学结果有重要影响。在比较使用不同技术和试剂的文献中的研究时，也必须谨慎。
Angiogenesis, or neovascularization, is known to play an important role in the neoplastic progression leading to metastasis. CD31 or Factor VIII-related antigen (F VIII RAg) immunohistochemistry is widely used in experimental studies for quantifying tumor neovascularization in immunocompromised animal models implanted with transformed human cell lines. Quantification, however, can be affected by variations in the methodology used to measure vascularization including antibody selection, antigen retrieval (AR) pretreatment, and evaluation techniques. To examine this further, we investigated the microvessel density (MVD) and the intensity of microvascular staining among five different human tumor xenografts and a mouse syngeneic tumor using anti-CD31 and F VIII RAg immunohistochemical staining. Different AR methods also were evaluated. Maximal retrieval of CD31 was achieved using 0.5 M Tris (pH 10) buffer, while maximum retrieval of F VIII RAg was achieved using 0.05% pepsin treatment of tissue sections. For each optimized retrieval condition, anti-CD31 highlighted small vessels better than F VIII RAg. Furthermore, the MVD of CD31 was significantly greater than that of F VIII RAg decorated vessels (p < 0.001). The choice of antibody and AR method has a significant affect on immunohistochemical findings when studying angiogenesis. One also must use caution when comparing studies in the literature that use different techniques and reagents.