| Project/Area Number |
03670776
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Nagoya University |
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Principal Investigator |
KIKKAWA Fumitaka Nagoya University School of Medicine Assistant Professor, 医学部, 講師 (40224985)
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| Co-Investigator(Kenkyū-buntansha) |
AOYAMA Toshifumi Shinshu Univ. Sch. Medicine Dept Biochem. Associate Prof., 医学部, 助教授 (50231105)
KAWAI Michiyasu Nagoya Univ. Sch. Medicine Dept obsgy Assist. Prof., 医学部, 助手 (10234014)
加納 武夫 名古屋大学, 医学部, 講師 (50169596)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Gynecologic Carcinoma / Carcinogenesis / Cytochrome P450 / RT-PCR / チトクロームP450 / チトクロ-ムP450 / PCR |
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| Research Abstract |
Most carcinogens are bioactivated by cytochrome P450(CYPs) and these enzymes within target cells are closely related to susceptibility to cancer. Since extrahepatic CYPs are typically at much lower levels, the existence and the role of CYP in extrahepatic tissues have been difficult to assess. In this study, we modified the reverse transcriprtase-ploymerase chain reaction (RT-PCR) to evaluate the quantities of CYP 2E1 mRNA in human endometrium. Total RNA from human endometrium was reverse transcribed and coamplified by PCR in the same tube containing both primer pairs of CYP 2E1 and beta-actin. The CYP 2E1 and beta-actin PCR products were 298 bp and 600 bp, respectively. The restriction enzyme mboi digested these two products to the predicted size for DNA fragments, demonstrating that both PCR products were specific and CYP 2E1 mRNA exists in human endometrium. CYP 1A1 mRNA was also examined, m but could not be detected clearly. Adding [alpha-^<32>P]dCTP to reaction mixture made it possible to quantify the relative yield of the CYP 2E1 PCR product in comparison with the beta-actin product. The ratio of the yield of the CYP 2E1 PCR product comparison with the beta-actin PCR product could be calculated at a point of 25 cycles of amplification. This ratio and serum estradiol levels were correlated positively (gamma=0.654, P<0.05), but not relationship to serum progesterone Levels was observed.
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