ESTABLISHMENT OF AN INTRA-OPERATIVE TECHNIQUE FOE DETECTION OF MICROMETASTASES IN GASTRIC CANCER SURGERY USING LAMP GENE AMPLIFICATION.
| Project/Area Number |
15591377
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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 | Chiba University |
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Principal Investigator |
HAYASHI Hedeki CHIBA UNIVERSITY, RESEARCH CENTER FOR FORTIER MEDICAL ENGINEERING, ASSOCIATE PROFESSOR, フロンティアメディカル工学研究開発センター, 助教授 (20312960)
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| Co-Investigator(Kenkyū-buntansha) |
OCHIAI Takenori CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, PROFESSOR, 大学院・医学研究院, 教授 (80114255)
GUNJI Yoshio CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 大学院・医学研究院, 助教授 (60241957)
SHIMADA Hideaki CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, ASSISTANT PROFESSOR, 大学院・医学研究院, 講師 (20292691)
SEKI Naohiko CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 大学院・医学研究院, 助教授 (50345013)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | GASTRIC CANCER / MICROMETASTASIS / SENTINEL LYMPH NODE / LAMP AMPLIFICATION / cDNA MICROARRAY / 微小転移診断 |
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| Research Abstract |
The expression profiles in gastric cancer cells obtained from surgical dissection of 20 gastric adenocarcinoma specimens were compared with those in the corresponding non-cancerous mucosa by cDNA microarray analysis. On the basis of sequence information, 4,608 independent clones were selected and used to produce the cDNA microarray. Twenty six genes that were commonly up-regulated and 44 genes that were commonly down-regulated in cancerous tissues were identified. This analysis also showed that expression of a gene identified, S100A11, was associated with the development of lymph node metastases. Additional real-time PCR analysis of the other 6 gastric cancer specimens underpinned this association.
LAMP (loop-mediated isothermal amplification) technique is an emerging technology to detect gene expressions of targeted sequences, with high sensitivity and specificity. Primers of this LAMP reaction targeting cytokeratin 19(CK19) gene were designed and applicability of micrometastases detec
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tion using the LAMP technique to a gastric cancer surgery was tested. Expression of CK19 gene could be detected in all of the total RNA prepared from 3x10^6 of MKN1,MKN28,MKN45,KATO-III, and SH10-TC, gastric cancer cell lines. Sensitivity to detect CK19 gene was also tested using MKN1 cell line, and up to 2x10^1 cells could be identified in the mixture of 2x10^7 white blood cells from a healthy volunteer. This technique was also applied to clinical specimens. CK19 gene amplification was detected in both gastric cancer tissue and lymph nodes with histo-pathologically positive metastasis. CK19 gene amplification could not be identified from mesenteric lymph nodes from patients without any cancerous lesions. Furthermore, CK19 gene amplification could be detected from tissue lysate prepared from operative specimens of gastric cancer patients without extraction procedure of RNA, as well. Diagnosis of metastasis could be performed within 1 hour using the LAMP technique with the tissue lysate analysis. Less
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Report
(3 results)
Research Products
(6 results)
