| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
The purpose of this study is gene expression profiling analysis and corresponding sub-classification of fibroblastic cells within neoplastic and non-neoplastic diseased conditions.
(1) Immunohistochemical analysis using paraffin-embedded sections and monoclonal antibodies to CD34, vimentin(vim), podoplanin/D2-40(pod), cytokeratin 8(CK8), alpha-smooth muscle actin (ASMA), and high molecular weight caldesmon (HCD) was performed in gastric and colorectal neoplastic and non-neoplastic diseases. Seven kinds of fibroblastic cells were proposed; i) CD34+vim+pod-CKS-ASMA-HCD-cells in normal tissue, ii) CD34-vim+pod+CK8-ASMA-HCD-cells in early phase of cancer invasion to submucosa, acute inflammation and recent localized circulatory disturbance, and post-irradiated tissue, iii) CD34-vim+pod+CK8-ASMA+HCD-cells in desmoplastic stroma of invasive adenocarcinomas and periglandular areas of gastrointestinal adenomas, iv) CD34-vim+pod-CK8-ASMA+HCD-cells in scar tissue, v) CD34-vim+pod+CK8+ASMA+HCD-cel
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ls seen in subserosal tissue involved by neoplastic and non-neoplastic diseases, vi) CD34-vim+pod-CK8-ASMA+HCD+ cells in normal colorectal crypts (pericryptal fibroblast), vii) CD34-vim+pod-CK8-ASMA+HCD- cells in peri-glandular areas of gastrointestinal adenomas.
(2) To confirm and/or revise the proposed subtypes of fibroblastic cells, cell isolation (fibroblastic cells, lymphocytes, etc) should be hurried from the fresh normal and diseased tissues, using cell type specific cell surface markers and magnetic beads (magnetic bead method). And then, both serial analysis of gene expression and DNA microarray analysis should be done.
(3) In performing the magnetic beads method, to choose good cell surface markers for fibroblasts is most important. Both normal fibroblasts and endothelial cells express CD34 in the cell surface, but epithelial cells, mature lymphocytes and other mesenchymal cells does not; normal fibroblast are detected by CD34 in a fraction which endothelial cells are removed by CD31 antibody.
(4) No specific cell surface markers exist for cancer-associated fibroblasts (CA-Fib). Most of the cancer-associated fibroblasts express membrane-truncated matrix metalloproteinase (MT1-MMP) in their cell membrane. Specific antibodies to MT1-MMP are suitable for CA-Fib. Less
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