ognizant Communication Corporation


VOLUME 12, NUMBER 1, 2000

Oncology Research, Volume 12, pp. 1-4, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Molecular Polarity in Endothelial Cells and Tumor-Induced Angiogenesis

Vincenzo Chiarugi, Marco Ruggiero, and Lucia Magnelli

Department of Pathology and Experimental Oncology, University of Firenze, Firenze, Italy

Endothelial cells expose receptors for vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) at the abluminal, basal surface that work as basic regulators of tumor-induced angiogenesis. Their specific localization makes them susceptible to the activity of tumor-released stimulatory factors, like VEGF/VPF, which induce proliferation of the endothelial cell toward the extracellular matrix. At the same time, VEGF/VPF stimulates endothelial cells to expose tissue factor (TF), the high-affinity transmembrane receptor and cofactor for cellular initiation of the plasma coagulation protease cascades through the extrinsic pathway, so generating thrombin. Thrombin exerts a number of activities: it forms an extracellular fibrin barrier from the VEGF/VPF-dependent fibrinogen extravasation; it activates progelatinase-A (proMMP-2), which destroys the basal membrane, allowing proliferation of endothelial cells (ECs) in the novel tumoral fibrin matrix; finally, it induces EC proliferation, potentiating the VEGF effect. Another important factor exposed at the abluminal endothelial cell surface is membrane type 1 matrix metalloproteinase (MT1-MMP), a membrane-bound metalloproteinase, which also activates progelatinase-A, allowing an alternative pathway to that of thrombin to destroy the basal membrane. In addition, we will see that MT1-MMP is also engaged in a direct, cell-associated fibrinolytic activity, essential for tubulogenesis of the novel outsprouting capillary.

Key words: Endothelial cell polarity; Angiogenesis; Clotting factors

Address correspondence to Prof. Vincenzo Chiarugi, Department of Pathology and Experimental Oncology, University of Firenze, Viale Morgagni 50.50134, Firenze, Italy. Tel: +39-0-55-411131; Fax: +39-0-55-416908; E-mail: Magnellil@USA.net

Oncology Research, Volume 12, pp. 5-10, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.
Mutational Alterations of the p16CDKN2A Tumor Suppressor Gene Have Low Incidence in Mesenchymal Chondrosarcoma

Dae Kyung Bae,1 Yong-Koo Park,2 Sung Gil Chi,2 Chun Woo Lee,1 and K. Krishnan Unni3

Departments of 1Orthopedic Surgery and 2Pathology, College of Medicine, Kyung Hee University, Seoul, Korea
3Department of Pathology, Mayo Clinic, Mayo Foundation, Rochester, MN

Mutational inactivation of the cyclin-dependent kinase inhibitors (CDKIs) (p16INK4A/MTS1) tumor suppressor gene has been found in a variety of human tumor types. To investigate the involvement of CDKI abnormality in mesenchymal chondrosarcoma, alterations of CDKIs were examined in human mesenchymal chondrosarcoma tissues using a quantitative DNA/PCR, PCR-SSCP. Seven of 33 specimens (21.2%) showed abnormally low levels of p16CDKN2A amplification, suggesting that the allelic deletion of the gene might be a less frequent event in progression of this tumor. To detect subtle sequence alterations such as point mutations, SSCP analysis of the entire coding region of the p16CDKN2A gene, exons 1, 2, and 3 regions, showed no altered SSCP patterns in 33 mesenchymal chondrosarcoma specimens. A low incidence of genetic alterations of the p16CDKN2A was found in mesenchymal chondrosarcoma. Through this study, we conclude that alteration of the p16CDKN2A gene does not participate significantly in the tumorigenesis of mesenchymal chondrosarcoma.

Key words: p16CDKN2A tumor suppressor gene; Mesenchymal chondrosarcoma

Address correspondence to Yong-Koo Park, M.D., Department of Pathology, Kyung Hee University Hospital, #1 Hoeki-Dong, Dongdaemoon-Ku Seoul, 130-702, Korea. Tel: ++82-2-958-8742; Fax: ++82-2-957-0489; E-mail: damia@chollian.net

Oncology Research, Volume 12, pp. 11-15, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Coamplification of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Genes in Methotrexate-Resistant Human Leukemia Cell Lines

Srinivasan Srimatkandada,1 Shyam K. Dube,2 Mark Carmen,1 and Joseph R. Bertino3

1Department of Medicine, Yale University School of Medicine, New Haven, CT 06510
2Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, College Park, MD 20742-4450
3Molecular Pharmacology and Therapeutics, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, NY 10021

Methotrexate (MTX)-resistant K562 human myelocytic leukemia sublines with 20- and 200-fold amplified dihydrofolate reductase (DHFR) genes localized to homogeneously staining regions (HSRs) on the long arms of chromosomes 5, 6, and 19 were used to examine whether other genes mapping close to the DHFR genes were coamplified. The gene for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, located on chromosome 5q13.3-14, was coamplified 4-14-fold, corresponding to the levels of resistance exhibited by these cells. Similar observations were made with a MTX-resistant subline of the promyelocytic leukemia cell line, HL-60R, with 200 gene copies of DHFR. These observations indicate a tight linkage of DHFR and HMG-CoA genes on chromosome 5q.

Key words: Methotrexate; Dihydrofolate reductase; Coamplification; Myelocytic leukemia cell lines

Address correspondence to Joseph R. Bertino, M.D., Molecular Pharmacology and Therapeutics, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021. Tel: (212) 639-8230; Fax: (212) 639-2767; E-mail: bertinoj@mskcc.org

Oncology Research, Volume 12, pp. 17-24, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

CD18/CD54(+CD102), CD2/CD58 Pathway-Independent Killing of Lymphokine-Activated Killer (LAK) Cells Against Glioblastoma Cell Lines T98G and U373MG

Fumio Komatsu and Michiko Kajiwara

Blood Transfusion Service, School of Medicine, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyoku, Tokyo 113-8519, Japan

For natural killer cell-mediated cytolysis (NK-lysis) and lymphokine-activated killer cell-mediated cytolysis (LAK-lysis), the co-stimulatory signals of CD18/CD54(+CD102) and CD2/CD58 pathways are essential. However, in this report, we describe a LAK-lysis that does not depend upon these two pathways. The killed cells were glioblastoma cell lines T98G and U373MG. The LAK cells were induced from peripheral blood lymphocytes in the presence of interleukin-2. 1) The T98G and U373MG did not express CD54 or CD102, but expressed CD58. 2) However, when they were pretreated with an anti-CD58 (TS2/9), the LAK-lysis was not blocked. 3) The LAK-lysis was markedly inhibited by pretreating with Concanamycin A and slightly inhibited by treating with antitumor necrosis factor-related apoptosis-inducing ligand (anti-TRAIL) antibody. 4) Nineteen percent of the LAK cells adhered to the T98G. The adhered LAK cells killed it. But nonadherent LAK cells could not kill the T98G or U373MG but killed lymphoblastoma cell lines Raji and NALM-6. These findings suggested that this type of the LAK-lysis might not depend upon the CD18/CD54(+CD102) pathway or CD2/CD58 pathway. The effector cells that killed the T98G and U373MG might not always be the same as the effector cells that killed the other cell lines. The LAK cells contain several subsets, and one of the subsets might kill these two target cell lines.

Key words: Lymphokine-activated killer cell; Adhesion molecules, Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL); T98G; U373MG; NK-92

Address correspondence to Fumio Komatsu, M.D., Blood Transfusion Service, School of Medicine, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyoku, Tokyo 113-8519, Japan. Tel: 81-3-5803-5646; Fax: 81-3-5803-5647.

Oncology Research, Volume 12, pp. 25-31, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.
Inhibitory Effect of Liposomal MDP-Lys on Lung Metastasis of Transplantable Osteosarcoma in Hamster

Yasuaki Nitta,1 Takashi Sugita,1 Yoshikazu Ikuta,1 and Teruo Murakami2

1Department of Orthopaedic Surgery, and 2Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine. Hiroshima 734-8551, Japan

MDP-Lys (N2-[(N-acetylmuramyl)-L-alanyl-D-isoglutaminyl]-N6-stearoyl-L-lysine), a macrophage activator, is a lipophilic derivative of muramyl dipeptide (MDP). Multilamellar liposome incorporated MDP-Lys was prepared using phosphatidylcholine and phosphatidylserine by conventional film method, and its inhibitory effect on lung metastasis was compared with MDP-Lys as a solution in hamster's osteosarcoma. The lung metastatic rates after transplantation of the tumor to a lower extremity, in which the extremity was amputated 3 weeks later, were 50% and 100% 3 and 7 weeks, respectively, after transplantation. The rates after amputation were reduced by the treatment with MDP-Lys proportionally to the logarithmic MDP-Lys dose, and the rates 7 weeks after transplantation were 55% and 60%, respectively, in the MDP-Lys solution (50 mg/day) and liposomal MDP-Lys (20 mg twice/week) groups. Fifty percent of hamsters treated with liposomal MDP-Lys survived for more than 6 months. Considering that hamsters had a lung metastasis rate of 50% before MDP-Lys treatment, liposomal MDP-Lys given at a dose of 20 mg twice /week was effective for inhibiting lung metastasis at a far lower dose of MDP-Lys than that given as a solution (40 mg vs. 350 mg per week). No significant side effect of liposomal MDP-Lys, as evaluated by the comparison of body weight changes among differently treated hamsters, was detected. This greater inhibitory effect of liposomal MDP-Lys can be considered to be due to the longer retention of the liposomal form in the lung.

Key words: Osteosarcoma; Lung metastasis; MDP-Lys; Liposome; Hamster

Address correspondence to Yasuaki Nitta, M.D., Department of Orthopaedic Surgery, Hiroshima University School of Medicine, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. Tel: 81-82-257-5233; Fax: 81-82-257-5234.

Oncology Research, Volume 12, pp. 33-41, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.
Angiogenic Interactions of Vascular Endothelial Growth Factor, of Thymidine Phosphorylase, and of p53 Protein Expression in Locally Advanced Gastric Cancer

A. Giatromanolaki,1,2 M. I. Koukourakis,1,3 G. P. Stathopoulos,5 A. Kapsoritakis,1 G. Paspatis,1,4 S. Kakolyris,3 E. Sivridis,1,2 V. Georgoulias,4 A. L. Harris,1,6 and K. C. Gatter1,6

1Tumour and Angiogenesis Research Group, 18 Dimokratias Avenue, Iraklion 71306, Crete, Greece
2Department of Pathology, Democritus University of Thrace, Greece
3Department of Radiotherapy and Oncology, Medical School, University of Thessalia, Larisa, Greece
4Department of Gastrenterology, Venizelion General Hospital, Crete, Greece
52nd Medical Division, Ippocration Hospital, University of Athens, Greece
6Departments of Cellular Science and ICRF Medical Oncology Unit, Oxford Radcliffe Hospital, Oxford, UK

The assessment of the angiogenic profile of tumors may become an important tool as a guide for the inclusion of novel drugs and molecular therapies into the standard chemoradiotherapy policy. Several studies have shown the prognostic importance of microvessel density (MVD) and of angiogenic factor expression in operable gastric cancer. In the present study we investigated, with immunohistochemistry the MVD, the expression of vascular endothelial growth factor (VEGF) and of thymidine phosphorylase (TP) expression, as well as the nuclear expression of p53 protein, in a series of patients with locally advanced inoperable gastric cancer. A strong association of VEGF with TP expression was noted (P = 0.005), and tumors coexpressing these factors had a statistically higher MVD (P = 0.0001). Nuclear p53 accumulation was also related to a high MVD (P = 0.004), and this was independent of VEGF or TP expression. Microvessel density showed a bell-shaped association with prognosis; cases with an intermediate MVD exhibit a favorable outcome (P < 0.05). A trend of nuclear TP expression to define a group of patients with poorer prognosis was noted (P = 0.06), while none of the remaining variables showed any significant association. The immunostaining results allowed the grouping of the angiogenic profile in four major categories: 1) highly vascularized tumors with VEGF and/or TP expression (about 36% of cases); 2) highly angiogenic tumors with p53 nuclear accumulation and low VEGF/TP expression (7% of cases); 3) poorly vascularized tumor with low VEGF/TP and negative nuclear p53 staining (32% of cases); 4) poorly vascularized tumors with TP expression (7% of cases). Specific therapies targeting hypoxia, VEGF, or TP expression as well as p53 gene therapy have entered clinical experimentation or are already available for clinical use. Using the suggested markers more than 80% of locally advanced gastric carcinomas can be grouped in different categories according to their angiogenic profile. Such a categorization may be useful for phase III trials on novel therapies targeting the major angiogenesis-related features studied here.

Key words: Gastric cancer; Angiogenesis; p53; VEGF; Thymidine phosphorylase; Platelet-derived endothelial cell growth factor (PD-ECGF)

Address correspondence to Dr. Alexandra Giatromanolaki, Tumour and Angiogenesis Research Group, 18 Dimokratias Avenue, Iraklion 71306, Crete, Greece. Tel: 081-284661; Fax: 081-284661; E-mail: targ@her.forthnet.gr

Oncology Research, Volume 12, pp. 43-49, 2000
0965-0407/00 $20.00 + .00
Copyright © 2000 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.
Expression of p53 and bcl-2 in Clinically Localized Prostate Cancer Before and After Neo-Adjuvant Hormonal Therapy

Anna M. Cesinaro,1 Mario Migaldi,1 Giovanni Ferrari,2 Giovanni Castagnetti,2 Alberto Dotti,2 Carmela De Gaetani,1 Paolo Ferrari,2 and Gian P. Trentini1

1Department of Morphological Sciences and Legal Medicine, Section of Anatomic Pathology, University of Modena and Reggio Emilia, Policlinico via del Pozzo 71, 41100 Modena, Italy
2Division of Urology-Azienda USL, Viale V.Veneto 9, 41100 Modena, Italy

The prognostic significance of p53 and bcl-2 expression in prostate carcinoma is currently under investigation. The aim of the present study was to analyze their expression in diagnostic biopsies and in prostatectomies performed after neo-adjuvant hormonal therapy to investigate their role in hormone resistance. One hundred and six patients with advanced prostate carcinoma were treated for 3 months with LHRH analogues before radical surgery. The expression of p53 and bcl-2 was analyzed by immunohistochemistry in all cases of prostatectomy and in available biopsies obtained before treatment, and was correlated with clinicopathologic parameters and follow-up. A significant increase in p53 expression was found following hormonal therapy, whereas no changes were observed in the expression of bcl-2. The increase in p53 did not correlate with the presence of therapy-induced morphological changes in prostate cancers, but it did correlate significantly with histologic grade and pathologic stage, biochemical progression of the disease, and short overall survival. At multivariate analysis, only grade and stage proved to be independent predictors of shorter survival. There were no correlations between bcl-2 and clinicopathologic variables whether in biopsies or in prostatectomies. The unfavorable clinical course associated with p53-positive carcinomas suggests that neo-adjuvant hormonal therapy may cause the selection of minor p53 mutated clones, rather than the induction of wild-type p53. In any case, the enhanced expression of p53 could label hormone-resistant cancers for further adjuvant therapy.

Key words: Prostate cancer; Hormonal therapy; p53; bcl-2; Hormone resistance; Prognosis

Address correspondence to Anna Maria Cesinaro, M.D., Department of Morphological Sciences and Legal Medicine, Section of Anatomic Pathology, University of Modena and Reggio Emilia, Policlinico via del Pozzo 71, 41100 Modena, Italy. Tel: 0039-059-424981/424808; Fax: 0039-059-424820; E-mail: cesinaro.a@policlinico.mo.it