1999 Fiscal Year Final Research Report Summary
Molecular cloning of highly expressed genes in renal cell carcinoma using cifferential display system
| Project/Area Number |
10671502
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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 |
Urology
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| Research Institution | AICHI MEDICAL UNIVERSITY |
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Principal Investigator |
YOSHIKAWA Kazuhiro AICHI MEDICAL UNIVERSITY, DEPT. OF PATHOLOGY , LECTURER, 医学部, 講師 (60109759)
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| Co-Investigator(Kenkyū-buntansha) |
TAKI Tomohiro AICHI MEDICAL UNIVERSITY, DEPT. OF PATHOLOGY , LECTURER, 医学部, 講師 (00288523)
MITSUI Kenji AICHI MEDICAL UNIVERSITY, DEPT. OF PATHOLOGY , LECTURER, 医学部, 講師 (20268014)
FUKATSU Hidetoshi AICHI MEDICAL UNIVERSITY, DEPT. OF PATHOLOGY , PROFESSOR, 医学部, 教授 (20065551)
SAGA Shinsuke AICHI MEDICAL UNIVERSITY, DEPT. OF PATHOLOGY , PROFESSOR, 医学部, 教授 (40144141)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Differential desplay / Thymosin beta-10 / Thymosin |
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| Research Abstract |
(Purposes) We had tried to isolate specifically and/or highly expressed genes in renal cell carcinoma (RCC). Using antibodies of the gene products, The detection of antigen from RCCs was investigated.
(Materials and Methods) Differential display was performed with total RNAs from renal cell carcinoma and normal kidney tissues. Unique genes expressed in RCC were cloned, sequences, searched homologous genes and analyzed mRNA expression in RCCs and normal kidneys. Using the peptide of Thymosin beta-10(17-43aa), which was coded by an isolated gene, we developed anti-Thymosin beta-10 antibody in Japanese white rabbit. Antibodies were specifically purified using affinity columns conjugated with Thymosin beta-10 Peptides(17-43aa and 36-43aa) and anti Thymosin beta-10 antibody(17-43aa) was conjugated with biotin. The reactivity of these antibodies were analyzed by ELISA using Thymosin beat-10 and Thymosin beta-4 peptides. Sandwich ELISA was performed to detect Thymosin beta-10 antigen in extrac
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tions from RCC tissues and cultured supernatants of RCCs, using anti Thymosin beta-10 antibody(17-43aa) as capture antibody and anti Thymosin beta-10 antibody(36-43aa) as tracer antibody.
(Results) Four cDNA clones were isolated by Differential display. These genes were highly expressed in renal cell carcinomas by Northern blot analysis. Two genes were homologous with Thymosin beta-10 and HLA-DR alpha chain, the others were unknown genes. We developed anti Thymosin beta-10 antibody using Thymosin beta-10 peptide(17-43aa) and purified using Thymosin beta-10(17-43aa and 36-43aa) conjugated affinity columns. Anti Thymosin beta-10 antibody(36-43aa) was specifically reacted with Thymosin beta-10,but not with Thymosin beta-4. Thymosin beta-10 antigen was able to detected in the extraction of RCC tissues and cultured supernatants of RCCs by sandwich ELISA using anti Thymosin beta-10 antibodies.
(Discussion) Thymosin beta-10 might be a marker for several carcinomas, because we could detect the antigen using the antibodies. Less
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Research Products
(6 results)
