Abstract
Background/Purpose
The microvessel density (MVD) of most malignant tumors is considered to be strongly related to metastasis and prognosis. Weidner’s “hot spot method” for determining MVD is in general use, but it is possible that cells other than endothelial cells will also be stained. In our previous study, no correlations were observed between MVD determined by the “hot spot method” and prognosis/metastasis. But, using the “lumen method,” we found a correlation with the number of vessel structures only. In the present study, we analyzed the staining of microvessels in pancreatic cancer, using light microscopy, confocal laser scan microscopy (CLSM), and transmission electron microscopy (TEM).
Methods
Microvessel staining of pancreatic cancer with CD34, factor VIII, and CD45 antibodies was examined in consecutive slices by light microscopy. For CLSM, freshly resected specimens were immunostained with factor VIII and fluorescein isothiocynate. For TEM, specimens were fixed with 2.5% glutaraldehyde, treated with 1% osmium tetroxide, and embedded in epoxy resin.
Results
Staining of vessels with CD34 and factor VIII antibodies appeared similar under light microscopy. However, CD34-stained consecutive slices were judged not to reveal vessel structures, and some cells stained with CD45 antibody were similar in appearance to CD34-stained cells. Under CLSM, irregular arrangements of neovascularization, consisting of many branches, were observed, but many positively stained cells not identified as vessels were also seen. Microvessels were distinctly identified under TEM, but the types of individual cells could not be determined.
Conclusions
An integrated, reproducible method for the measurement of MVD is vital. For pancreatic cancer, the “lumen method” is recommended.
Similar content being viewed by others
References
Weidner N, Semple J, Welich W, Folkman J. Tumor angiogenesis and metastasis: correlation in invasive breast carcinoma. N Engl J Med 1991;324:1–8.
Viecent-Salomon A, Carton M, Zafrani B, Freneaux P, Nicolas A, Massemin B, et al. Long term outcome of small size invasive breast carcinomas independent from angiogenesis in a series of 685 cases. Cancer 2001;92:249–256.
Lee JS, Kim HS, Jung JJ, Kim YB, Park CS, Lee MC. Correlation between angiogenesis, apoptosis and cell proliferation in invasive ductal carcinoma of the breast and their relation to tumor behavior. Anal Quant Cytol Histol 2001;23:161–168.
Gunel N, Akcali Z, Coskum U, Akyol G, Yamac D, Yenidunya S. Prognostic importance of tumor angiogenesis in breast carcinoma with adjuvant chemotherapy. Pathol Res Pract 2002;198:7–12.
Guidi AJ, Berry DA, Broadwater G, Helmchen B, Bleiweiss IJ, Budman DR, et al. Association of angiogenesis and disease outcome in node-positive breast cancer patients treated with adjuvant cyclophosphamide, doxorubicin, and fluorouracil: a Cancer and Leukemia Group B Correlative Science Study From Protocols 8541/8869. J Clin Oncol 2002;20:732–742.
Bossi P, Viale G, Lee A, Alfano RM, Coggi G, Bosari S. Angiogenesis in colorectal tumors: microvessel quantitation in adenomas and carcinomas with clinicopathological correlations. Cancer Res 1995;55:5049–5053.
Fox SH, Whalen GF, Sanders MM, Burleson JA, Jennings K, Kurtzman S, et al. Angiogenesis in normal tissue adjacent to colon cancer. J Surg Oncol 1998;69:230–234.
Weidner N, Folkman J. Tumor vascularity as a prognostic factor in cancer. Adv Oncol 1996, 167–190.
Vermeulen PB, Gasparini G, Fox SB, Toi M, Martin L, McCulloch P, et al. Quantification of angiogenesis in solid human tumors: an international consensus on the methodology and criteria of evaluation. Eur J Cancer 1996;32:2474–2484.
Uzzan B, Nicolas P, Cucherat M, Perret GY. Microvessel density as a prognostic factor in women with breast cancer: a systematic review of the literature and meta-analysis. Cancer Res 2004;64:2941–2955.
Hall NR, Fish DE, Hunt N, Goldin RD, Guillou PJ, Monson RT. Is the relationship between angiogenesis and metastasis in breast cancer real? Surg Oncol 1992;1:223–229.
Takagi K, Takada T, Amano H. A high peripheral microvessel density count correlates with a poor prognosis in pancreatic cancer. J Gastroenterol 2005;40:402–408.
Folkman J. Tumor angiogenesis: therapeutic implications. N Engl J Med 1971;285:1182–1186.
Folkman J. Clinical applications of research on angiogenesis. N Engl J Med 1995;333:1757–1763.
Ellis LM, Takahashi Y, Fenoglio CJ, Cleary KR, Bucana CD, Evans DB. Vessel counts and vascular endothelial growth factor expression in pancreatic adenocarcinoma. Eur J Cancer 1998;34:337–340.
Karademir S, Sokmen S, Terzi C, Sagol O, Ozer E, Astarcioglu H, et al. Tumor angiogenesis as a prognostic predictor in pancreatic cancer. J Hepatobiliary Pancreat Surg 2000;7:489–495.
Fujioka S, Yoshida K, Yanagisawa S, Kawakami M, Aoki T, Yamazaki Y. Angiogenesis in pancreatic carcinoma. Cancer 2001;92:1788–1797.
Niedergethmann M, Hildenbrand R, Wostbrock B, Hartel M, Sturm JW, Richter A, et al. High expression of vascular endothelial growth factor predicts early recurrence and poor prognosis after curative resection for ductal adenocarcinoma of the pancreas. Pancreas 2002;25:122–129.
Fujimoto K, Hosotani R, Wada M, Lee JU, Koshiba T, Miyamoto Y, et al. Expression of two angiogenic factors, vascular endothelial growth factor and platelet-derived endothelial cell growth factor in human pancreatic cancer, and its relationship to angiogenesis. Eur J Cancer 1998;34:1439–1447.
Ikeda N, Adachi M, Taki T, Huang C, Hashida H, Takabayashi A, et al. Prognostic significance of angiogenesis in human pancreatic cancer. Br J Cancer 1999;79:1553–1563.
Nagakawa Y, Aoki T, Kasuya K, Tsuchida A, Koyanagi Y. Histologic features of venous invasion, expression of vascular endothelial growth factor and matrix metalloproteinase-2 and matrix metalloproteinase-9, and the relation with liver metastasis in pancreatic cancer. Pancreas 2002;24:169–178.
Ueda T, Oda T, Kinoshita T, Konishi M, Nakahashi C, Takahashi S, et al. Neovascularization in pancreatic ductal adenocarcinoma: microvessel count analysis, comparison with noncancerous regions and other types of carcinomas. Oncol Rep 2002;9:239–245.
Guo L, Burke P, Lo SH, Gandour-Edwards R, Lau D. Quantitative analysis of angiogenesis using confocal laser scanning microscopy. Angiogenesis 2001;4:187–191.
Isner J, Asahara T. Angiogenesis and vasculogenesis as therapeutic strategies for postnatal neovascularization. J Clin Invest 1999;103:1231–1236.
Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, Silver M, et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 1999;85:221–228.
Author information
Authors and Affiliations
About this article
Cite this article
Takagi, K., Takada, T., Amano, H. et al. Analysis of microvessels in pancreatic cancer: by light microscopy, confocal laser scan microscopy, and electron microscopy. J Hepatobiliary Pancreat Surg 15, 384–390 (2008). https://doi.org/10.1007/s00534-007-1241-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00534-007-1241-6