The elucidation of denelopment and progression of the scirrhous gastric cancer by the new technology to amplify an extremely small amount od RNA
| Project/Area Number |
13671284
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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 |
Digestive surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
UEDA Tetsuya the University of Tokyo, Faculty of Medicim, research associate, 医学部附属病院, 助手 (40272562)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Hiroki Genetics Division, National Cancer Center Research Institute, section head, 分子腫瘍学部, 室長(研究職) (60235265)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2001: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | scirrhous gastric cancer / micyoarray / LCM / microdissection / a small amount of RNA / TALPAT / peritoneal dissemination / スキルス胃がん |
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| Research Abstract |
1. The technology to amplify an extremely small amount of RNA from about 100 gastric cancer cells collected by laser-captured microdissection is necessary for the microarray analysis of me poorly differentiated gastric cancer like the scirrhous gastric cancer. We have developed and optimized the method called TALPAT which combined adaptor ligation-mediated PCR and T7 RNA polymerase-mediated RNA amplification. Total RNA from the 100 cells of gastric cancer cell line was amplified by TALPAT and analyzed with Affymetrix gene chip (12,629 genes), and very high reproducibility was shown. RNA from 100〜1000 cells collected by LCM from the cancer tissue was amplified by TALPAT, and it could be applied to the microarray analysis.
2. We analyzed the 40 gastric cancer tissues of various types of differentiation with Affymetrix gene chip, and compared gene expression profiles. As a result, it was proved that gastric cancer which showed purely the characteristics of die scirrhous gastric cancer coul
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d be divided from por2 of the un-differentiation type gastric cancer. We separated cancer cells and fibroblasts from that type of gastric cancer tissue by LCM, and amplified total RNA by TALPAT, and analyzed with Affymetrix gene chip. We obtained the expression profiles which were useful for the elucidation of development and progression of the scirrhous gastric cancer.
3. During the process of searching for the genes which were specific to the scirrhous gastric cancer, we obtained the 11 genes which could be applied to the way of diagnosing a minute peritoneal dissemination during the operation by RT-PCR of peritoneal washes. We examined the 108 RNA samples from peritoneal washes by RT-PCR which were amplified by TALPAT for the prevention of depletion. Five genes showed high specificity and they could be applied to qualitative PCR, but 6 genes showed a little low specificity, and it was considered to be necessary that they were applied quantitative PCR or tmmunocytochemistry. As for five genes, it was proved that not only peritoneal dissemination but also other types of recurrence could be predicted by RT-PCR. Less
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Report
(3 results)
Research Products
(3 results)
