Three-Dimensional (3D) Imaging of Tumor Angiogenesis and its Inhibition: Evaluation of Tumor Vascular-Targeting Agent Efficacy in the DMBA-induced Rat Breast Cancer Model by Confocal Laser Scanning Microscopy (CLSM)

There is considerable interest in tumor angiogenesis and its inhibition, because tumor angiogenesis plays an important role in tumor growth and distant metastasis, and is highly associated with overall survival and relapse free-survival in cancer patients. This study was performed to determine the mechanisms underlying tumor angiogenesis and its inhibition using the angiogenesis inhibitor TNP 470 and antimitotic drug, docetaxel (Taxotere). Variable 3D image analysis of tumor vasculature after chemotherapy was performed by CLSM using the DMBA-induced rat breast cancer model. 3D images of the vasculature in DMBA-induced rat breast cancer were characterized by increased microvessel density and anastomosing, irregularly branched microvessel networks. After administration of TNP-470 or docetaxel, tumor microvessel density was markedly decreased and tumor microvessel networks were markedly destroyed. Furthermore, tumor microvessels less than 30 μm in diameter were not found. The remaining tumor microvessels (30-60 μm in diameter) were mainly localized in the periphery of the tumor. These observations suggest that docetaxel might have the ability to inhibit tumor angiogenesis as well as TNP-470. CLSM with the FI method is a valuable technique for estimation of the efficacy of anti-angiogenic chemotherapy.

[1]  H. Dvorak,et al.  Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. , 1983, Science.

[2]  L. Orci,et al.  Potent synergism between vascular endothelial growth factor and basic fibroblast growth factor in the induction of angiogenesis in vitro. , 1992, Biochemical and biophysical research communications.

[3]  S. Ramakrishnan,et al.  Vascular permeability factor gene expression in normal and neoplastic human ovaries. , 1994, Cancer research.

[4]  S. Ramakrishnan,et al.  Inhibition of angiogenesis and tumour growth by VEGF121–toxin conjugate: differential effect on proliferating endothelial cells , 2000, British Journal of Cancer.

[5]  M A Konerding,et al.  Angiogenesis inhibitors endostatin or TNP-470 reduce intimal neovascularization and plaque growth in apolipoprotein E-deficient mice. , 1999, Circulation.

[6]  M. Shimamura,et al.  Powerful antiangiogenic activity of herbimycin A (named angiostatic antibiotic). , 1989, The Journal of antibiotics.

[7]  J. Folkman.  The influence of angiogenesis research on management of patients with breast cancer , 2004, Breast Cancer Research and Treatment.

[8]  Y. Uehara,et al.  The microvasculature of the 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumour , 1991, Virchows Archiv A.

[9]  Y. Naito,et al.  Paclitaxel-induced remission in docetaxel-refractory anthracycline-pretreated metastatic breast cancer , 2000, Anti-cancer drugs.

[10]  E. Manseau,et al.  Increased expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in kidney and bladder carcinomas. , 1993, The American journal of pathology.

[11]  M. Zucchetti,et al.  Epidoxorubicin and docetaxel as first-line chemotherapy in patients with advanced breast cancer: a multicentric phase I-II study. , 2000, Annals of oncology : official journal of the European Society for Medical Oncology.

[12]  Y. Kakeji,et al.  Preclinical studies of the combination of angiogenic inhibitors with cytotoxic agents , 2004, Investigational New Drugs.

[13]  A. Harris,et al.  Novel growth regulatory factors and tumour angiogenesis. , 1991, European journal of cancer.

[14]  C. Verschraegen,et al.  Use of docetaxel (Taxotere) in patients with paclitaxel (Taxol) hypersensitivity. , 2000, Anti-cancer drugs.

[15]  Grietje Molema,et al.  Tumor Infarction in Mice by Antibody-Directed Targeting of Tissue Factor to Tumor Vasculature , 1997, Science.

[16]  D. Ingber,et al.  Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumour growth , 1990, Nature.

[17]  S. Ergün,et al.  CEA-related cell adhesion molecule 1: a potent angiogenic factor and a major effector of vascular endothelial growth factor. , 2000, Molecular cell.

[18]  D. Chaplin,et al.  Antivascular approaches to solid tumour therapy: evaluation of tubulin binding agents. , 1996, The British journal of cancer. Supplement.

[19]  R. Osamura,et al.  Dynamic MR imaging and tumor angiogenesis in DMBA-induced rat breast cancer: Three dimensional (3D) reconstructed image analysis of tumor microvessels by confocal laser scanning microscopy , 1997, Breast cancer.

[20]  J. Itoh,et al.  Vascular networks and endothelial cells in the rat experimental pituitary glands and in the human pituitary adenomas , 2003, Microscopy research and technique.

[21]  D. Ingber,et al.  Cytostatic inhibition of endothelial cell growth by the angiogenesis inhibitor TNP-470 (AGM-1470). , 1994, British Journal of Cancer.

[22]  P. Chiarugi,et al.  Endostatin: A Promising Drug for Antiangiogenic Therapy , 1999, The International journal of biological markers.

[23]  Georg Breier,et al.  Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo , 1992, Nature.

[24]  W. Fan,et al.  In vitro evaluation of schedule-dependent interactions between docetaxel and doxorubicin against human breast and ovarian cancer cells. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[25]  W. Gradishar,et al.  Clinical trials of antiangiogenic agents , 1997, Current opinion in oncology.

[26]  D. Chaplin,et al.  Tumour vasculature as a target for cancer therapy. , 1999, British journal of cancer.

[27]  A. Bikfalvi,et al.  Target molecules for anti-angiogenic therapy: from basic research to clinical trials. , 2000, Critical reviews in oncology/hematology.

[28]  R. D'Amato,et al.  Inhibition of angiogenesis and breast cancer in mice by the microtubule inhibitors 2-methoxyestradiol and taxol. , 1997, Cancer research.

[29]  J. Itoh,et al.  Three-dimensional imaging of tumor angiogenesis. , 2000, Analytical and quantitative cytology and histology.

[30]  J. Folkman,et al.  Efficacy of antiangiogenic therapy with TNP-470 in superficial and invasive bladder cancer models in mice. , 2000, Urology.

[31]  D. Collins,et al.  Inhibition of angiogenesis by suramin. , 1992, Cancer research.

[32]  J. Folkman.  What is the evidence that tumors are angiogenesis dependent? , 1990, Journal of the National Cancer Institute.

[33]  L. Milas,et al.  Lack of correlation between mitotic arrest or apoptosis and antitumor effect of docetaxel , 1999, Cancer Chemotherapy and Pharmacology.

[34]  R. Kerbel,et al.  'Accidental' anti-angiogenic drugs. anti-oncogene directed signal transduction inhibitors and conventional chemotherapeutic agents as examples. , 2000, European journal of cancer.

[35]  D. Ingber,et al.  Angiostatic steroids. Method of discovery and mechanism of action. , 1987, Annals of surgery.

[36]  T. Imazawa,et al.  Antitumor effect of the angiogenesis inhibitor agm-1470 and its combination effect with tamoxifen in dmba induced mammary-tumors in rats. , 1993, International journal of oncology.

[37]  Eric C. Griffith,et al.  Cell cycle inhibition by the anti-angiogenic agent TNP-470 is mediated by p53 and p21WAF1/CIP1. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Ducreux,et al.  Phase I and pharmacokinetic study of docetaxel and irinotecan in patients with advanced solid tumors. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[39]  J. Ascherman,et al.  Docetaxel (taxotere) extravasation: a report of five cases with treatment recommendations. , 2000, Annals of plastic surgery.

[40]  P. Nakane.  RECENT PROGRESS IN THE PEROXIDASE‐LABELED ANTIBODY METHOD * , 1975, Annals of the New York Academy of Sciences.

[41]  D. Chaplin,et al.  Anti—vascular approaches to solid tumour therapy: Evaluation of vinblastine and flavone acetic acid , 1995, International journal of cancer.

[42]  A. Hara,et al.  Inhibitory effect of TNP-470 on hepatic metastasis of mouse neuroblastoma. , 2000, The Journal of surgical research.

[43]  William Arbuthnot Sir Lane,et al.  Endostatin: An Endogenous Inhibitor of Angiogenesis and Tumor Growth , 1997, Cell.

[44]  J. Itoh,et al.  A New Approach to Three-dimensional Reconstructed Imaging of Hormone-secreting Cells and Their Microvessel Environments in Rat Pituitary Glands by Confocal Laser Scanning Microscopy , 2000, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[45]  A. Matsuno,et al.  Confocal laser scanning microscopic imaging of subcellular organelles, mRNA, protein products, and the microvessel environment , 2001 .

[46]  H. Dvorak,et al.  Vascular permeability factor (vascular endothelial growth factor) gene is expressed differentially in normal tissues, macrophages, and tumors. , 1992, Molecular biology of the cell.