2001 Fiscal Year Final Research Report Summary
Analysis of gene expression associated with inflammatory bowel disease using cDNA microarray
| Project/Area Number |
12470249
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | Tohoku University |
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Principal Investigator |
SASAKI Iwao Tohoku Univ., Graduate School of Med., Dept. of Surg., Professor, 大学院・医学系研究科, 教授 (60125557)
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| Co-Investigator(Kenkyū-buntansha) |
FUNAYAMA Yuji University Hospital of Tohoku Univ., Lecturer, 医学部附属病院, 講師 (50192315)
FUKUSHIMA Kouhei Tohoku Univ., Graduate School of Med., Dept of Surg., Research Associate, 大学院・医学系研究科, 助手 (20271900)
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| Project Period (FY) |
2000 – 2002
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| Keywords | intestinal flora / epithelial cells / inflammatory bowel disease / CRP-ductin / microarray / ulcerative colitis / Crohn's disease |
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| Research Abstract |
Summary of murine epithelial gene screening by cDNA microarray : Poly (A) RNA was extracted from total RNA of isolated epithelial cells. Epithelial gene expression of the colon and small intestine was compared among Specific pathogen-free, Germ-free and 3 day bacteria reconstituted mice by cDNA microarray (GEM microarray, Incyte Genomics Co.). This system is able to compare expression of 8000 genes between two groups. Differential expression of reg IIIβ and Illγ was also detected by microarray, demonstrating that both differential display and microarray are appropriate, sensitive, reproducible and complementary methods. We selected 62 genes based on differential expression and high expression level. Those include 43 known genes and 19 EST (expression sequence tag). Forty-two clones have been used as probes in northern blotting and 20 not yet. In 42 clones, differential expression was confirmed in 35 clones and 5 not. Two clones were not evaluated due to low expression of the genes. RNAs from DSS-colitis were also used in some experiments.
Epithelial induction of a human homologue of murine CRP-ductin. Deleted in Malignant Brain Tumors 1 (DMBT1) : Induction of CRP-ductin mRNA was initially detected by cDNA microarray. This gene was also induced in DSS-colitis even after recovery from active inflammation by the removal of DSS. Northern blotting revealed that the human homologue, DMBT1 with 7 kb in size, was weakly expressed in control but remarkably induced in two-thirds of IBD isolates. An additional 6.5 kb band was noticed in one-third of IBD isolates. Real-time RT-PCR demonstrated that DMBT1mRNA level was significantly higher in both UC and CD.
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[Publications] Ogawa H., Fukushima K., Naito H., Funayama Y., Unno M., Takahashi K., Kitayama T., Matsuno S., Ohtani H., Takasawa S., Okamoto H., Sasaki I.: "Increased expression of HIP/PAP and Regenerating Gene III in Human Inflammatory Bowel Disease and a Murine Bacterial Reconstitution Model"Inflammatory Bowel Diseases. 9(3). 162-170 (2003)
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「研究成果報告書概要(欧文)」より
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