Establishment of detection system of micrometastasis in the regional lymph nodes of colorectal cancer by RT-PCR for matrilysin

基质溶解素RT-PCR检测结直肠癌区域淋巴结微转移体系的建立

• 批准号: 10671207
• 负责人: ICHIKAWA Yasushi
• 金额: $ 2.43万
• 依托单位: YOKOHAMA CITY UNIVERSITY, SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671207/
• 关键词: matrilysin; micrometastasis; lymphnode metastasis; GFP; 血管新生抑制; KDR; Flk-1; RT-PCR; 微小転移診断

Detection system of micrometastasis in the regional lymph nodes of colorectal cancer utilizing RT-PCR for matrilysin (MTR) was established. MTR method could detect as small as 10^4 cancer cells in one lymph node. Sensitivity of MTR method was significantly higher than conventional histopathologic examination using H-E staining. About 60 % of cases showing conventional method negative, and MTR method positive, changed into H-E staining positive by additional sequential sectioning, however, no cases showing negative in the both methods changed into positive in the same examination. Moreover, MTR method was significantly sensitive than the other famous molecular biological method detecting micrometastasis using PCR for point mutated k-ras. We are keeping watch on prognosis of cases showing n0 by conventional method and positive by MTR method and no recurrence has been detected in those cases. In our department, radical lymph node dissection was performed for patients of colorectal cancer, then local recurrence or recurrence of regional lymph nodes was signifcantly less than the other facilities. The other problem is false positive of PCR because of its super-high sensitivity. Then we are now trying to perform quantitative PCR using TaqMan PCR method for matrilysin. To establish this new technique, further examination is necessary.

建立了RT-PCR检测结直肠癌区域淋巴结微转移的检测体系。MTR法可在一个淋巴结内检出小至104个癌细胞。MTR法的敏感性明显高于常规HE染色组织病理学检查。常规方法为阴性，MTR法为阳性的病例中，约有60%的病例经附加序贯切片后转为H-E染色阳性，但在同一检查中，两种方法均未发现阴性病例转为阳性。此外，MTR法检测k-ras基因点突变的微转移敏感性明显高于其他著名的分子生物学方法。我们正在密切关注常规方法显示N0和MTR方法阳性的病例的预后，这些病例尚未发现复发。我科对结直肠癌患者行根治性淋巴清扫术，局部复发率或局部复发率明显低于其他术式。另一个问题是，由于聚合酶链式反应的超高敏感性而导致的假阳性。然后，我们现在正在尝试使用TaqMan聚合酶链式反应方法对基质溶素进行定量聚合酶链式反应。为了建立这项新技术，有必要进行进一步的检查。

项目成果

Ichikawa Y, Ishikawa T, Momiyama N, Yamaguchi S, Masui H, Hasegawa S, Chishima T, Takimoto A, Kitamura H, Akitaya T, Hosokawa T, Mitsuhashi M, Shimada H.: "Detection of regional lymph node metastases in colon cancer by using RT-PCR for matrix metalloprote

Ichikawa Y、Ishikawa T、Momiyama N、Yamaguchi S、Masui H、Hasekawa S、Chishima T、Takimoto A、Kitamura H、Akitaya T、Hosokawa T、Mitsuhashi M、Shimada H.：“结肠癌区域淋巴结转移的检测

Ichikawa Y. et al.: "Detection of regional lymph node metastases in colon cancer by using RT-PCR for matrix metalloproteinase 7, matrilysin"Clinical and Experimental Metastasis. 16. 3-8 (1998)

Ichikawa Y. 等人：“通过使用 RT-PCR 检测基质金属蛋白酶 7、基质溶解素来检测结肠癌区域淋巴结转移”临床和实验转移。

Ichikawa Y. et al.: "MMP-7 (matrilysin) accelerated growth of human umbilical vein endothelial cells"Cancer Letter. 8. 95-100 (2002)

Ichikawa Y.等人：“MMP-7（基质溶解素）加速人脐静脉内皮细胞的生长”癌症信。

Yasushi Ichikawa: "Detection of Regionl Lymph Node Metastasis in Colon Caacer by RT-PCR for Matrily" Clinical and Experimental Metastasis. 16. 3-8 (1998)

Yasushi Ichikawa：“通过 RT-PCR 检测结肠癌区域淋巴结转移”临床和实验转移。

ICHIKAWA.Y et al.: "MMP-7(matrilysin) accelerated growth of human umbilical vein endothelial cells"Cancer Letter. 177. 95-100 (2002)

ICHIKAWA.Y 等人：“MMP-7（基质溶解素）加速人脐静脉内皮细胞的生长”癌症信。