| Project/Area Number |
14570809
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | KAWASAKI MEDICAL SCHOOL |
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Principal Investigator |
FUJIMOTO Wataru KAWASAKI MEDICAL SCHOOL, MEDICAL SCHOOL, PROFESSOR, 医学部, 教授 (50165429)
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| Co-Investigator(Kenkyū-buntansha) |
UCHIDA Takafumi KAWASAKI MEDICAL SCHOOL, MEDICAL SCHOOL, ASSISTANT, 医学部, 助手 (50368635)
大森 謙太郎 川崎医科大学, 医学部, 講師 (20278930)
幸田 衞 川崎医科大学, 医学部, 助教授 (50153474)
山口 雅英 川崎医科大学, 医学部, 講師 (60299196)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2004: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | glucocorticoid receptor / real-time RCR / autoimmune bullous disease / real-time RT-PCR |
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| Research Abstract |
Alternative splicing of the human glucocorticoid receptor (hGR) pre-mRNA generates two highly homologous isoforms, reserved hGR α and hGR β. In contrast to hGR α, hGRβ does not bind glucocorticoids and is transcriptionally inactive. Recently, hGRβ was suggested to play a role in glucocorticoid resistance because the expression of hGR β is increased in peripheral blood mononuclear cells (PBMCs) in patients with fatal asthma and glucocorticoid-resistant ulcerative colitis. Our previous study demonstrate that the expression of hGR β is significantly lower tha that of hGR α in PBMC from normal volunteers as revealed by semiquantitative RT-PCR and immunohistochemistry. The aim of this study was to clarify whether analysis of hGR α and hGR β can predict the response to glucocorticoids in patients with autoimmune bullous diseases. Total RNA obtained from peripheral blood mononuclear cells (PBMCs) of 5 patients with pemphigus or bullous pemphigoid and 7 healthy volunteers was analysed by semiquantitative RT-PCR and one-step RT-PCR using specific TaqMan probe for hGR α and β. Semiquantitative RT-PCR revealed that the expression of hGR β is significantly lower than that of hGR α in PBMC both in patients with BP and healthy volunteers. However, in contrast to the results obtained from semiquantitative RT-PCR, the real time PCR could not demonstrate quantitative difference of expression level of both hGR α and hGR β. The second experiment with various concentration of primers specific for both hGR α and hGR β showed that relative ratio of hGR α and hGR β mRNA expression could fluctuate according to the primer concentration. The third experiment using another new primers and a TaqMan probe specific for hGR α and hGR β also failed to establish the optimal conditions for quantitative analysis of hGR α and hGR β.
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