2002 Fiscal Year Final Research Report Summary
THE ANALYSIS OF CHROMOSOMAL GAINS AND LOSSES IN PRIMARY MALIGNANTMELANOMA-COMPARISON WITH SPITZ NEVUS-
| Project/Area Number |
13670875
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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 | SHINSHU UNIVERSITY |
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Principal Investigator |
MATSUMOTO Kazuhiko SHINSHU UNIVERSITY HOSPITAL, ASSISTANT PROFESSOR, 医学部附属病院, 講師 (40165882)
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| Project Period (FY) |
2001 – 2002
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| Keywords | MELANOMA / SPITZ NEVUS / MICRODISSECIION / COMPARATIVE GENOMIC HYBRIDIZATION |
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| Research Abstract |
SPITZ NEVUS IS A BENIGN NEOPLASM OF MELANOCYTES THAT CAN BE DIFFICULT OR IMPOSSIBLE TO DISTINGUISH FROM MELANOMA BY CLINICAL AND HISTOPATHOLOGIC EXAMINATION. WE TRIED TO FIND CHROMOSOMAL GAIN AND LOSSES BY COMPARATIVE GENOMIC HYBRIDIZATION (CGH) IN FORMALIN-FIXED, PARAFFIN ENMBEDDED TISSUES FROM MELANOMA, AND COMPARE WITH THAT FROM SPITZ NEVUS. HOWEVER, THE SIZE OF DNA EXTRACTED FROM FORMARIN-FIXED, PARAFFIN-EMBEDDED TISSUE IS TOO SMALL FOR CGH. THEREFORE, WE FIRST TRIED TO ESTABLISH THE CGH METHOD USING MELANOMA CELL LINE, MMG1. REFERENCE DNA WAS EXTRATED FROM NORMAL FEMALE PERIPHERAL BLOOD LYMPHOCYTES. WE GOT THE FOLLOWING RESULTS.
1. EXTRACTION OF DNA : DNA EXTRACTOR WB KIT (WAKO, OSAKA) CAN BE USED FOR THE EXTRACTION FROM THE CELL LINES.
2. LABELING TIME : ONE AND A HALF HOUR FOR MMG1 CELLS. THREE HOURS FOR PERIPHERAL BLOOD LYMPHOCYTES.
3. PRECIPITATION TIME WITH ETHANOL : -80℃, ONE HOUR
4. HYBRIDIZATION TIME: 37℃, 3 DAYS
WE HAVE ESTABLISHED THE CGH METHOD IN OUR LABORATORY THE FINAL PROBLEM IS THE DNA EXTRACTION FROM FORMARIN-FIXED, PARAFFIN-EMBEDDED TISSUES. WE WILL SOLVE THE PROBLEM SOON AND WE CAN FIND CHROMOSOMAL GAIN AND LOSSES BY COMPARATIVE GENOMIC HYBRIDIZATION (CGH) IN FORMALIN-FIXED, PARAFFIN ENMBEDDED TISSUES FROM MELANOMA AND SPITZ NEVUS.
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