Clarification of the lymph node metastasis mechanism by orthotopic implantation models and application to inhibition of the metastasis

原位种植模型阐明淋巴结转移机制及其在抑制转移中的应用

• 批准号: 17591422
• 负责人: YAMAGUCHI Koji
• 金额: $ 1.73万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591422/
• 关键词: gastric cancer; orthotopic implantation; VEGF-C gene; lymph node metastasis; cDNA microarray

To investigate the molecular mechanisms by which carcinoma cells metastasize, we have established liver metastatic models. Human gastric carcinoma cell line AZ521 (5 × 10^6/0.1ml) was transplanted orthotopically into the stomach of nude mice using cell suspension technique. Athymic female BALB/c nu/nu mice, 6-7 weeks old, weighing 18-22g were used. After from 6 to 8 weeks, the liver and regional lymph nodes with a few metastatic foci of AZ521 cells were dissected, we established AZH1G and AZL5G. The same procedure was repeated using AZH1G and AZL1G, and AZH5G and AZL5G was established upon the fifth cycle of stepwise selection. We have also established AZ3P3G using intraperitoneally and orthotopically stepwise selection methods. 1) AZL5G produced lymph node mestastasis in 85.0%, and MKNL3G produced lymph node mestastasis in 60.0% of mice. 2) Each metastatic cell grew significantly larger than parental AZ521 in vivo. 3) A cell motility assay demonstrated that the migration ability of each metastatic cell was clearly higher than that of parental cell lines. 4) The expression of several cell adhesion molecules by FACS analysis showed that the incidence of α_1 and α_2 integrins expression in AZL5G cells was significantly higher than in AZ521. Then, MKNL3G cells expressed suggestive up-regulation of α_2 integrin. Moreover, Comprehensive analysis of the biological behavior of highly metastatic cell lines and the less metastatic same parental lines were demonstrated the differences in adhesive activity and VEGF production. Up-regulated genes were endothelin-A receptor and TGF-beta II R alpha in AZL5G. Above all, our experimental models should be useful for investigation of the mechanisms of metastasis in human gastric carcinoma. Up-regulated genes in MKNL3G and inhibitory methods are currently underway.

为了研究癌细胞转移的分子机制，我们建立了肝转移模型。采用细胞悬液法将人胃癌细胞株AZ 521（5 × 10^6/0.1ml）原位移植于裸鼠胃内。使用无胸腺雌性BALB/c nu/nu小鼠，6-7周龄，重18- 22 g。6 ~ 8周后，切除肝脏和有少量转移灶的区域淋巴结，建立AZH 1G和AZL 5G细胞株。使用AZH 1G和AZL 1G重复相同的程序，并在第五轮逐步选择时建立AZH 5G和AZL 5G。我们还建立了AZ 3 P3 G使用腹腔内和原位逐步选择方法。1)AZL 5G的淋巴结转移率为85.0%，MKNL 3G的淋巴结转移率为60.0%。2)每个转移细胞在体内生长显著大于亲本AZ 521。3)细胞运动性测定表明，每个转移细胞的迁移能力明显高于亲本细胞系。4)流式细胞仪检测AZL 5G细胞粘附分子的表达，结果显示AZL 5G细胞中整合素α 1和α 2的表达率明显高于AZ 521细胞。MKNL 3G细胞表达α_2整合素。此外，综合分析高转移细胞系和低转移细胞系的生物学行为，证实了粘附活性和VEGF产生的差异。AZL 5G表达上调的基因为内皮素A受体和TGF-β II R α。本实验模型的建立为进一步研究胃癌转移机制奠定了基础。MKNL 3G中上调的基因和抑制方法目前正在进行中。

项目成果

Plasma level of a 5-fluorouracil metabolite, fluoro-beta-alanine correlates with dihydropyrimidine dehydrogenase activity of peripheral blood mononuclear cells in 5-fluorouracil treated patients.

在接受 5-氟尿嘧啶治疗的患者中，5-氟尿嘧啶代谢物氟-β-丙氨酸的血浆水平与外周血单核细胞的二氢嘧啶脱氢酶活性相关。

• 发表时间: 2006
• 期刊: J Exp Clin Cancer Res. 25
• 作者: Furuhata T;Okita K;Kimura Y;et al.
• 通讯作者: et al.

A novel isoform of TUCAN is overexpressed in human cancer tissues and suppresses both caspase-8-and caspase-9-mediated apoptosis

• DOI: 10.1158/0008-5472.can-04-4649
• 发表时间: 2005-10-01
• 期刊: CANCER RESEARCH
• 影响因子: 11.2
• 作者: Yamamoto, M;Torigoe, T;Sato, N
• 通讯作者: Sato, N

The interplay between gastric cancer cell lines and PBMCs mediated by the CC chemokine RANTES plays an important role in tumor progression.

由 CC 趋化因子 RANTES 介导的胃癌细胞系和 PBMC 之间的相互作用在肿瘤进展中发挥着重要作用。

• 发表时间: 2005
• 期刊: J Exp Clin Cancer Res 24(3)
• 作者: Okita K;Furuhata T;Kimura Y;et al.
• 通讯作者: et al.