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A subject, compound technology, applied in the direction of DNA/RNA fragments, compound screening, antibody medical components, etc.
Inactive Publication Date: 2007-05-30
ONCOTHERAPY SCI INC
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[0014] It has been repeatedly reported that peptide-stimulated peripheral blood mononuclear cells (PBMCs) from specific healthy donors produced significant levels of IFN-γ in response to the peptide, but hardly HLA-γ in the chromium-51 release assay. A24 or -A0201 restriction patterns exert cytotoxic effects against tumor cells (Kawano et al., Cancer Res 60:3550-8 (2000); Nishizaka et al., Cancer Res 60:4830-7 (2000); Tamura et al., Jpn J Cancer Res 92:762-7(2001))
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Embodiment 1
[0219] 1. General method
[0220] Patient and Tissue Samples
[0221] Tissue samples were obtained with informed consent from 26 cancer patients undergoing radical prostatectomy. All surgical samples were in T2a-T3a clinical stage with or without N1, with a Gleason score of 5-9. Histopathological blood diagnosis was performed by a pathologist using LMM. All samples were buried in TissueTek OCT medium (Sakura, Tokyo, Japan) immediately after surgical resection, and stored at -80°C for future use. Among the 26 PRC tissues, 20 cancers and 10 advanced PINs had sufficient RNA quantity and quality for microarray studies.
[0222] Laser microbeam microdissection and T7-based RNA amplification
[0223] LMM and T7-based RNA amplification were performed as previously described. Prostate tumor cells and normal prostate ductal epithelial cells were selectively isolated using the EZ cutting system (SL Microtest GmbH, Germany) with a pulsed ultraviolet narrowbeam-focus laser accord...
Embodiment 2
[0248] 1. General method
[0249] Cell Lines and Tissue Samples
[0250] Human pancreatic cell lines PK45P, KLM1 and MIA-PaCa2 (ATCC number: CRL-1420) were obtained from Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku University. Such cells are publicly available.
[0251] Isolation of Overexpressed Genes in PDACa Cells Using cDNA Microarray
[0252] Fabrication of cDNA microarray slides has been described (Ono K, Tanaka T, Tsunoda T, Kitahara O, Kihara C, Okamoto A, Ochiai K, Takagi T, and Nakamura Y. Cancer Res., 60:5007-5011, 2000) . For each analysis of expression profiling, cDNA microarray slides containing approximately 27000 DNA spots were prepared in duplicate. Briefly, total RNA was purified from PDACa cells, and normal pancreatic ductal epithelium was obtained by microdissection from 18 pancreatic carcinoma tissues. RNA suitable for microarray experiments was obtained based on performing T7 RNA amplification. Ali...
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Abstract
Objective methods for diagnosing a predisposition to developing prostate cancer (PRC) are described herein. In one embodiment, the diagnostic method involves the determining a expression level of EphA4. The present invention further provides methods of screening for therapeutic agents useful in the treatment of PRC, methods of treating PRC. The invention also features a method for inhibiting growth of a cancer cell by contacting the cell with a composition of a siRNA of EPHA4. Methods of treating cancer are also within the invention. The invention also features products, including nucleic acid sequences and vectors as well as to compositions comprising them, useful in the provided methods. The invention also provides a method for inhibiting of tumor cell, for example pancreatic cancer cell, particularly pancreatic ductal adenocarcinoma (PDACa).
Description
[0001] This application claims priority to US Provisional Application Serial No. 60 / 548,335, filed February 27, 2004, and US Provisional Application Serial No. 60 / 555,809, filed March 24, 2004, the entire contents of which are incorporated herein by reference. technical field [0002] The present invention relates to methods of detecting and diagnosing a predisposition to prostate cancer (PRC) and pancreatic ductal adenocarcinoma (PDACa). The present invention also relates to methods of treating and preventing prostate cancer and pancreatic ductal adenocarcinoma (PDACa), in particular the present invention relates to EphA4. Background technique [0003] Prostate cancer (PRC) is one of the most common malignancies in men and the second leading cause of cancer-related death in the United States and Europe (Gronberg et al., 2003). Serum prostate-specific antigen (PSA) detection detects early PRC and is now the gold standard for screening for PRC in high-risk individuals. [00...
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