ESTABLISHMENT OF AN INTRA-OPERATIVE TECHNIQUE FOE DETECTION OF MICROMETASTASES IN GASTRIC CANCER SURGERY USING LAMP GENE AMPLIFICATION.

建立使用灯基因扩增检测胃癌手术中微转移的术中技术。

• 批准号: 15591377
• 负责人: HAYASHI Hedeki
• 金额: $ 2.24万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591377/
• 关键词: GASTRIC CANCER; MICROMETASTASIS; SENTINEL LYMPH NODE; LAMP AMPLIFICATION; cDNA MICROARRAY; 微小転移診断

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 … More 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

通过cDNA微阵列分析，将20个胃腺癌标本的手术解剖获得的胃癌细胞中的表达谱与相应的非癌粘膜中的表达谱进行比较。根据序列信息，选择了 4,608 个独立克隆并用于产生 cDNA 微阵列。确定了癌组织中 26 个通常上调的基因和 44 个通常下调的基因。该分析还表明，所鉴定的基因 S100A11 的表达与淋巴结转移的发生有关。对其他 6 个胃癌标本的额外实时 PCR 分析也支持了这种关联。LAMP（环介导等温扩增）技术是一种检测目标序列基因表达的新兴技术，具有高灵敏度和特异性。设计了针对细胞角蛋白 19 (CK19) 基因的 LAMP 反应引物，并测试了使用 LAMP 技术在胃癌手术中检测微转移的适用性。 MKN1、MKN28、MKN45、KATO-III和SH10-TC胃癌细胞系3x10^6制备的总RNA中均可检测到CK19基因的表达。还使用 MKN1 细胞系测试了检测 CK19 基因的灵敏度，在健康志愿者的 2x10^7 个白细胞混合物中最多可以识别出 2x10^1 个细胞。该技术也应用于临床标本。在胃癌组织和组织病理学阳性转移的淋巴结中均检测到CK19基因扩增。在没有任何癌性病变的患者的肠系膜淋巴结中无法鉴定出 CK19 基因扩增。此外，无需提取RNA，也可以从胃癌患者手术标本制备的组织裂解液中检测到CK19基因扩增。使用 LAMP 技术结合组织裂解物分析，可以在 1 小时内完成转移诊断。较少的

项目成果

Detection of sentinel lymph nodes in gastric cancers based on Immunohistochemical analysis of micrometastases

基于微转移免疫组织化学分析的胃癌前哨淋巴结检测

• 发表时间: 2004
• 期刊: Journal of Surgical Oncology 87
• 作者: Mikito Mori;Tomoaki Karube
• 通讯作者: Tomoaki Karube

S100A11 gene identifies by in-house cDNA microarray as an accurate predictor of lymph node metastases of gastric cancer

S100A11 基因通过内部 cDNA 微阵列鉴定为胃癌淋巴结转移的准确预测因子

• 发表时间: 2004
• 期刊: Oncology Reports 11
• 作者: Mikito Mori

癌治療の新たなる試み 新編III,センチネルナビゲーション手術

癌症治疗的新尝试，新版三，哨兵导航手术

• 发表时间: 2005
• 作者: Mikito Mori;Tomoaki Karube;Mikito Mori;西條 長宏 編
• 通讯作者: 西條 長宏 編

林 秀樹, 落合武徳: "Sentinel lymph node navigation surgery-センチネルリンパ節:現況と将来-手術への応用 胃癌"Current Therapy. 21・7. 31-35 (2003)

Hideki Hayashi、Takenori Ochiai：“前哨淋巴结导航手术-前哨淋巴结：现状和未来-在胃癌手术中的应用”21・7（2003）。

S100A11 gene identified by in-house cDNA microarray as an accurate predictor of lymph node metastases of gastric cancer.

• DOI: 10.3892/or.11.6.1287
• 发表时间: 2004-06
• 期刊: Oncology reports
• 影响因子: 4.2
• 作者: M. Mori;H. Shimada;Y. Gunji;H. Matsubara;H. Hayashi;Y. Nimura;Masaki Kato;M. Takiguchi;T. Ochi