Loges Sonja, Clausen Henning, Reichelt Uta, Bubenheim Michael, Erbersdobler Andreas, Schurr Paulus, Yekebas Emre, Schuch Gunter, Izbicki Jakob, Pantel Klaus, Bokemeyer Carsten, Fiedler Walter
Department of Internal Medicine II, University Hospital Hamburg-Eppendorf, Hamburg, Germany.
Clin Cancer Res. 2007 Jan 1;13(1):76-80. doi: 10.1158/1078-0432.CCR-06-1324.
Angiogenesis and lymphangiogenesis are important steps in tumor growth and dissemination and are of prognostic importance in solid tumors. The determination of microvessel density (MVD) by immunohistology is subject to considerable variability between different laboratories and observers. We compared MVD determination by immunohistology and quantitative real-time PCR and correlated the results with clinical variables.
The expression of endothelial antigens vascular endothelial cadherin (CD144), P1H12 (CD146), tie-2, and VEGFR-2, and lymphatic endothelial markers VEGFR-3, Prox, and LYVE was assessed by quantitative PCR (qPCR) in primary surgical samples. The expression of angiogenetic growth factors VEGF-A, VEGF-C, VEGF-D, angiopoietin-1, and angiopoietin-2 was quantified by PCR and correlated with MVD and clinical variables.
The expression of endothelial antigens vascular endothelial cadherin (CD144), P1H12 (CD146), tie-2, and VEGFR-2 correlated with each other in 54 samples of primary esophageal cancer (P < 0.0001 for all comparisons). MVD determined immunohistologically by CD31 staining in a subgroup of 35 patients correlated significantly with the qPCR method. The expression of angiogenetic growth factors VEGF-A, VEGF-C, VEGF-D, angiopoietin-1, and angiopoietin-2 was significantly associated with MVD (P < 0.0001 for all comparisons). Analysis of the expression of lymphendothelial markers VEGFR-3, Prox, and LYVE revealed concordant results, indicating that quantification of lymphendothelial cells is possible by qPCR. The presence of lymph node metastasis on surgical specimens was significantly correlated with MVD (P < 0.003), VEGFR-2 (P < 0.048), and VEGF-C (P < 0.042) expression.
These results indicate that quantification of MVD by qPCR in surgical samples of esophageal carcinoma yields similar results with immunohistology. Interestingly, the extent of angiogenesis and lymphangiogenesis was not related in individual tumor samples. Lymph node metastases could be predicted by MVD and VEGF-C expression.
血管生成和淋巴管生成是肿瘤生长和扩散的重要步骤，对实体瘤的预后具有重要意义。通过免疫组织化学法测定微血管密度（MVD）在不同实验室和观察者之间存在相当大的差异。我们比较了免疫组织化学法和定量实时PCR法测定MVD，并将结果与临床变量相关联。
通过定量PCR（qPCR）评估原发性手术样本中内皮抗原血管内皮钙黏蛋白（CD144）、P1H12（CD146）、tie-2和VEGFR-2以及淋巴管内皮标志物VEGFR-3、Prox和LYVE的表达。通过PCR对血管生成生长因子VEGF-A、VEGF-C、VEGF-D、血管生成素-1和血管生成素-2的表达进行定量，并与MVD和临床变量相关联。
在54例原发性食管癌样本中，内皮抗原血管内皮钙黏蛋白（CD144）、P1H12（CD146）、tie-2和VEGFR-2的表达相互关联（所有比较P<0.0001）。在35例患者的亚组中，通过CD31染色免疫组织化学测定的MVD与qPCR方法显著相关。血管生成生长因子VEGF-A、VEGF-C、VEGF-D、血管生成素-1和血管生成素-2的表达与MVD显著相关（所有比较P<0.0001）。对淋巴管内皮标志物VEGFR-3、Prox和LYVE表达的分析显示结果一致，表明通过qPCR可以对淋巴管内皮细胞进行定量。手术标本上淋巴结转移的存在与MVD（P<0.003）、VEGFR-2（P<0.048）和VEGF-C（P<0.042）表达显著相关。
这些结果表明，通过qPCR对食管癌手术样本中的MVD进行定量与免疫组织化学法得到的结果相似。有趣的是，在单个肿瘤样本中血管生成和淋巴管生成的程度无关。淋巴结转移可以通过MVD和VEGF-C表达来预测。
