|ognizant Communication Corporation|
Featuring PRECLINICAL AND CLINICAL CANCER THERAPEUTICS
VOLUME 17, NUMBER 5
Oncology Research, Vol. 17, pp. 193-203
0965-0407/08 $90.00 + .00
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.
In Vitro Antioxidative Potential of Lactoferrin and Black Tea Polyphenols and Protective Effects In Vivo on Carcinogen Activation, DNA Damage, Proliferation, Invasion, and Angiogenesis During Experimental Oral Carcinogenesis
P. Vidjaya Letchoumy,1 K. V. P. Chandra Mohan,1 J. J. Stegeman,2 H. V. Gelboin,3 Y. Hara,4 and S. Nagini1
1Department of Biochemistry and Biotechnology,
Faculty of Science, Annamalai University, Tamil Nadu, India
2Woods Hole Oceanographic Institute, Woods Hole, MA, USA
3National Cancer Institute, Bethesda, MD, USA
4Mitsui Norin Co. Ltd., Shizuoka, Japan
The present study was designed to evaluate the in vitro antioxidant potential of bovine lactoferrin (bLF) and black tea polyphenols [Polyphenon-B (P-B)] as well as in vivo inhibitory effects on the development of 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinomas. Antioxidant activity was screened using a panel of assays including 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2´-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), hydroxyl radical anion (OH.), superoxide anion (O2.-), and nitric oxide (NO) radical scavenging assays as well as assay for reducing power. The chemopreventive potential of bLF and P-B was assessed in the HBP model based on the modulatory effects on DMBAinduced oxidative DNA damage as well as the expression of proteins associated with carcinogen activation (CYP1A1, CYP1B1), cell proliferation [cyclin D1, proliferating cell nuclear antigen (PCNA), glutathione S-transferase pi (GST-P)], angiogenesis [vascular endothelial growth factor (VEGF), VEGF receptor 1 (VEGFR1)], and invasion and metastasis [matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of MMP-2 (TIMP-2)]. Both bLF and P-B showed high radical scavenging activity and reductive potential. Although administration of bLF and P-B alone suppressed DMBA-induced HBP tumors, combined administration of bLF and P-B was more effective in inhibiting HBP carcinogenesis by inhibiting oxidative DNA damage, carcinogen activation, cell proliferation, invasion, and angiogenesis. Our study suggests that the antioxidative property of bLF and P-B may be responsible for chemoprevention of HBP carcinogenesis by modulating multiple molecular targets.
Key words: Antioxidants; Chemoprevention; Carcinogen activation; Cell proliferation; Angiogenesis
Address correspondence to Dr. S. Nagini, Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamil Nadu, India. Tel: +91-4144-239842; Fax: +91-4144-238145/238080; E-mail: email@example.com or firstname.lastname@example.org
Maximizing Immune Responses: The Effects of Covalent Peptide Linkage to Beta-2-Microglobulin
Stephen M. Bauer,1 Marc A. Williams,1,2,3 Alan P. Howell,4 Edward Schwarz,5 Ernest S. Smith,4 and Maurice Zauderer4
1Division of Pulmonary and Critical
Care Medicine, University of Rochester School of Medicine, Rochester, NY,
2Department of Environmental Medicine, University of Rochester, Rochester, NY, USA
3Lung Biology and Disease Program, University of Rochester School of Medicine, Rochester, NY, USA
4Vaccinex, Inc., Rochester, NY, USA
5Department of Pathology and Laboratory Medicine, University of Rochester, Rochester, NY, USA
Major histocompatability molecules (MHC) are involved in presentation of peptide antigens for recognition by the immune system. The density and stability of presented peptides is a critical parameter in determining the magnitude of the immune response. Increasing the half-life and density of an MHC class I-peptide complex should promote a stronger cytotoxic T lymphocyte (CTL) response to clinically important peptides, including those that exhibit low or suboptimal MHC class I binding affinity. We hypothesized that the covalent linkage of a known tumor antigen peptide to beta-2-microglobulin (b2m) would increase peptide immunogenicity and, therefore, in vivo effectiveness as an antitumor vaccine in BALB/c mice. The iL3 peptide fusion protein (iL3-L12-hb2m) was developed based on the mutant iL3 peptide, derived from the L3 ribosomal protein, and expressed in the mutagenized murine fibroblastic tumor cell line, BCA34. The iL3-L12-b2m and a negative control fusion protein utilizing the H-2Kd-restricted NP(147-155) influenza peptide (NPL12-hb2m) were both produced in E. coli for exogenous antigen presentation by dendritic cells. In vitro, the iL3-L12-hb2m protein was found to stabilize H-2Kd over time on the surface of H-2Kd-expressing target cells and sensitized them to peptide-specific CTL-mediated lysis. Furthermore, mice immunized with dendritic cells pulsed with the iL3-L12-hb2m protein rejected a challenge with BCA34 cells significantly more so than mice immunized with dendritic cells pulsed with free peptide and hb2m. We conclude that vaccines incorporating peptides covalently linked to b2m may have future potential in the specific targeting of humanmalignancy.
Key words: Fusion proteins; Cancer; Dendritic cells
Address correspondence to Stephen M. Bauer, Ph.D., Division of Pulmonary and Critical Care Medicine, University of Rochester School of Medicine, 601 Elmwood Avenue, Box 692, Rochester, NY 14642-8692, USA. Tel: 585-276-3270; E-mail: Stephen_Bauer@urmc.rochester.edu or Maurice Zauderer, Ph.D., President and CEO, Vaccinex, Inc., 1895 Mount Hope Avenue, Rochester, NY 14620, USA. Tel: 585-271-2700; E-mail:email@example.com
Recombinant Human Monoclonal IgA Antibody Against CEA to Recruit Neutrophils to CEA-Expressing Cells
Jun Zhao,1 Motomu Kuroki,1,2 Hirotomo Shibaguchi,1,2 Lin Wang,2 Qun Huo,2 Noboru Takami,3 Toshihiro Tanaka,1,2 Tetsushi Kinugasa,1 and Masahide Kuroki1,2
1Molecular Oncology Center, Faculty
of Medicine, Fukuoka University, Fukuoka 814-0180, Japan
2Department of Biochemistry, Faculty of Medicine, Fukuoka University, Fukuoka 814-0180, Japan
3Radioisotope Laboratory, Faculty of Medicine, Fukuoka University, Fukuoka 814-0180, Japan
IgA is able to trigger antibody-dependent cellular cytotoxicity (ADCC) by recruiting neutrophils expressing the Fc receptor for the CHa chain. We herein describe the preparation of a human recombinant anti-CEA IgA antibody to kill carcinoembryonic antigen (CEA)-expressing tumor cells via ADCC by neutrophils. A single chain Fv (scFv) gene was constructed using a cDNA library of a hybridoma clone that produces a human anti-CEA monoclonal IgG4 (C2-45). The scFv gene, linked with a CHa gene, was inserted into the pBK283 vector, which was cotransfected into BmN4 insect cells with the wild-virus BmNPV. After cloning and amplification, the recombinated virus was injected into silkworm larvae. The resulting human recombinant IgA, designated as 45scFvLCHa, was purified from hemolymph by Ni-affinity chromatography and characterized by ELISA, Western blotting, and the ADCC assay. 45scFvLCHa with an IgA antigenicity was bound to CEA and showed effective killing of the CEA-expressing cells in the presence of IFN-g-activated neutrophils. These data suggest the recombinant anti-CEA IgA antibody recruiting neutrophils maybe a useful means for the antibody-based immunotherapy of human CEA-expressing tumors.
Key words: Carcinoembryonic antigen (CEA); IgA; Neutrophils; Antibody-dependent cellular cytotoxicity (ADCC); Immunotherapy
Address correspondence to Dr. Motomu Kuroki, Ph.D., Department of Biochemistry, Faculty of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan-ku, Fukuoka, 814-0180, Japan. Tel: (+81) 92 801 1011, Ext. 4367; Fax: (+81) 92 801 3600; E-mail: firstname.lastname@example.org
Synergistic Effect of 5-Aza-2´-Deoxycytidine and Genistein in Combination Against Leukemia
Noël J.-M. Raynal,1,2 Michel Charbonneau,1 Louise F. Momparler,2 and Richard L. Momparler2
1INRS-Institut Armand-Frappier, Université
du Québec, Laval, Quebec, H7V 1B7, Canada
2Service of Hematology/Oncology, Centre de recherche, Ste-Justine Hospital, Dept. de pharmacologie, Université de Montréal, Montreal, Quebec, H3T 1C5, Canada
5-Aza-2´-deoxycytidine (5-AZA-CdR), a potent inhibitor of DNA methylation, is an effective agent for the treatment of leukemia. The aim of this study was to investigate the antileukemic activity of this epigenetic agent in combination with genistein, a nontoxic isoflavone with chemopreventive activity. The combined treatment produced a synergistic loss of clonogenicity in human myeloid (HL-60) and lymphoid (MOLT-3) leukemic cell lines. Genistein alone showed a significant antileukemic activity against murine 5-AZA-CdRresistant cells, and this effect was enhanced when used in combination with 5-AZA-CdR. The combined treatment also produced a synergistic increase in life span of mice with L1210 leukemia. These results suggest that genistein may have the potential to increase the clinical efficacy of 5-AZA-CdR for the treatment of leukemia.
Key words: 5-Aza-2´-deoxycytidine; Epigenetic; Leukemia; Genistein; Chemotherapy; Drug resistance
Address correspondence to Richard L. Momparler, Service of Hematology/Oncology, Centre de recherche, Ste-Justine Hospital, Université de Montréal, 3175 Cote Ste. Catherine Road, Montreal, Quebec, H3T 1C5, Canada. Tel: +514 342 4931, ext 6140; Fax: +514 345 4801; E-mail: email@example.com
Follicle Stimulating Hormone Receptor in Ovarian Surface Epithelium and Epithelial Ovarian Cancer
Chinmoy K. Bose
Institut für Reproduktionsmedizin, 48149 Münster, Germany
Epithelial ovarian cancer (EOC) has remained an enigmatic disease, the etiology of which is mostly unknown. Considering the age incidence around menopause, a "gonadotrophin theory" has been proposed, and considering the association with infertility, an "incessant ovulation" theory has been proposed. EOC originates from ovarian surface epithelium (OSE), which not only secretes cytokines/growth factors and steroids but expresses gonadotrophin and steroid hormone receptors as well. The most important gonadotrophin receptor is the follicle stimulating hormone receptor (FSHR), which has definite oncogenic potential and is a probable candidate for oncogenesis. In this article, we review existing knowledge of FSHR in ovary, in OSE, and in epithelial ovarian cancer and try to establish relative importance of this receptor over its ligand. A systematic review through PubMed was done on the subject of FSHR and its metabolism. For the obvious difficulty of meager amounts of tissue available, most of studies so far see it in granulosa cells and cell lines rather than in OSE. Thus, effort was made to deduce workable knowledge that can establish its role and can then be applied in OSE and EOC. There is great deal of information regarding metabolism of FSHR, including regulation of its gene expression, isoforms, and pathways of desensitization and degradation with which ovarian cancer etiology researchers have to be familiar with, and there are a number of steps where manipulation may stop carcinogenesis. Hormone therapy of such cancer has so far been only mildly active, probably because we do not understand the role and mechanism of action of FSH and FSHR in the hypothalamo-pituitary-ovarian axis in development of such cancer.
Key words: Follicle stimulating hormone receptor (FSHR); Ovarian surface epithelium (OSE); Epithelial ovarian cancer (EOC)
Address correspondence to Dr. Chinmoy K. Bose at his current address: 8D, Mathur Sen Garden Lane, Kolkata 700006, India. Fax: 00913328430741; E-mail: firstname.lastname@example.org