ASSOCIATION OF GLUCOCORTICOID RECEPTOR β EXPRESSION AND STEROID INSENSITIVITY IN PATIENTS WITH AUTOIMMUNE BULLOUS DISEASES
| Project/Area Number |
14570809
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Dermatology
|
|---|
| Research Institution | KAWASAKI MEDICAL SCHOOL |
|---|
Principal Investigator |
FUJIMOTO Wataru KAWASAKI MEDICAL SCHOOL, MEDICAL SCHOOL, PROFESSOR, 医学部, 教授 (50165429)
|
|---|
| 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)
|
|---|
| Project Period (FY) |
2002 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| 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)
|
|---|
| Keywords | glucocorticoid receptor / real-time RCR / autoimmune bullous disease / real-time RT-PCR |
|---|
| 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.
|
Report
(4 results)
Research Products
(8 results)
