| Project/Area Number |
07457386
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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 |
Obstetrics and gynecology
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| Research Institution | Kansai Medical University |
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Principal Investigator |
KANZAKI Hideharu Kansai Medical University, Professor, 医学部, 教授 (80135566)
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| Co-Investigator(Kenkyū-buntansha) |
YASUDA Katsuhiko Kansai Medical University,, 医学部, 講師 (90174507)
FUJITA Jun Kyoto University, Clinical Molecular Biology, Professor, 医学部, 教授 (50173430)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1996: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1995: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Human Endometrium / Implantation / cDNA Subtraction / Progesterone / Tissue Transglutaminase / TIMP-3 |
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| Research Abstract |
It is generally believed that the cellular events during the establishment and maintenance of pregnancy are mediated through the expression of specific steroid-regulated genes in the uterus. The identification of these steroid-regulated genes is crucial for a clear understanding of the molecular and cellular processes that control uterine endometrial growth and differentiation during pregnancy. In the present study, we have tried to identify the genes involved in decidualization, and then investigated the role of the molecules coded by the genes in stromal cell differentiation and implantation process. By employing subtractive hybridization, we have constructed cDNA library enriched with progesterone dependent genes. While analyzing about 2,000 clones derived from this subtracted cDNA library, we have identified different 12 genes whose expression were up-regulated with progesterone. Kinetic analysis of 3 clones showed that this subtracted cDNA library may contain genes involved in var
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ious step of progesterone-induced decidualization. One of these clones was identical to the human tissue inhibitor of metalloproteinase-3 (TIMP-3), a recently identified member of the human TIMP family. Induction of TIMP-3 mRNA expression was evident when human endometrial stromal cells were cultured with progesterone for 6 days. Other 2 genes were induced as early as 2-6 hours after progesterone stimulation ; one of them was revealed to be tissue transglutaminase type II (TGase) and the other was an unreported gene. In situ hybridization revealed the expression of TIMP-3 mRNA in predecidualized stromal cells and decidual cells of human endometria. Because the role of MMP and their specific inhibitors, TIMP,in the process of tissue remodeling has been demonstrated in various tissues, TIMP-3 may be important for the controlled invasion of fetal trophoblasts at the feto-maternal interface. On the other hand tissue TGase was important in the process of decidualization since the inhibition of the enzyme activity by specific inhibitor of antisense oligonucleotide, the in vitro differentiation of endometrial stromal cells was inhibited. Immunohistochemical studies suggested the importance of TGase in human endometrial differentiation toward blastocyst implantation, and Northern blotting for the unreported gene (UK10) showed that the gene expression was significantly decreased in patients of implantation failure diagnosed by repeated IVF-ET treatment. Less
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