Establishment and Characterization of Highly Metastatic Human Gastric Carcinoma Cell Line Established by Orthotopic Tumor Cell Implantation

原位肿瘤细胞植入高转移性人胃癌细胞系的建立和表征

• 批准号: 12671240
• 负责人: YAMAGUCHI Koji
• 金额: $ 1.15万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671240/
• 关键词: orthotopic implantatiopn; ymph node metastasis; liver metastasis; peritoneal dissemination; gastric carcinoma; 同所性移植; 胃癌細胞株; ヌードマウス; 接着分子

To investigate the molecular mechanisms by which carcinoma cells metastasize, we have established liver metastatic models. Human gastric carcinoma cell line AZ521 (5 X 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 AZH5G produced multiple round liver mestastasis in 87.5%, and AZ3P3G developed peritoneal metastasis in 80.0% 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 AZ521. 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. AZH5G cells expressed suggestive up-regulation of α_1, α_2, α_3 and α_5 integrins, and AZ3P3G cells expressed up-regulation ofα_2,α_3,α_5 andα_6 integrins. Moreover, Comprehensive analysis of the biological behavior of highly metastatic cell lines and the less metastatic same parental line were demonstrated the differences in adhesive activity and VEGF production. Above all, our experimental models should be useful for investigation of the mechanisms of metastasis in human gastric carcinoma.

为了研究癌细胞转移的分子机制，我们建立了肝转移模型。采用细胞悬浮法将人胃癌细胞株AZ521(5×10~(-6)/0.1ml)原位移植到裸鼠的胃内。选用6-7周龄、体重18-22g的雌性BALB/c-nu/nu裸鼠。6~8周后，解剖有少量AZ521细胞转移灶的肝脏和区域淋巴结，建立AZH1G和AZL5G。用AZH1G和AZL1G重复相同的步骤，在第五轮逐步选择时建立AZH5G和AZL5G。我们还利用腹膜内和原位逐步选择的方法建立了AZ3P3G。1)AZL5G有85.0%的小鼠发生淋巴结转移，AZH5G有87.5%的小鼠发生多发圆形肝转移，AZ3P3G有80.0%的小鼠发生腹膜转移。2)体内各转移细胞生长均明显大于亲本AZ521。3)细胞运动实验表明，各转移细胞的迁移能力均明显高于AZ521。4)流式细胞仪分析多种细胞黏附分子的表达情况，发现α_1和α_2整合素在AZL5G细胞中的表达明显高于AZ521细胞。AZH5G细胞表达α_1、α_2、α_3和α_5整合素，AZ3P3G细胞表达α_2、α_3、α_5和α_6整合素表达上调。此外，对高转移细胞系和低转移亲本细胞系的生物学行为的综合分析表明，高转移细胞系和低转移亲本细胞系的黏附活性和血管内皮生长因子的产生存在差异。最重要的是，我们的实验模型对于研究人胃癌的转移机制是有用的。

项目成果

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Yamaguchi K.: "Establishment and Characterization of a Human Gastric Carcinoma Cell Line that is Highly Metastatic to Lymph Nodes"J Ext Clin Cancer Res.. 19(1). 113-120 (2000)

Yamaguchi K.：“高度转移至淋巴结的人胃癌细胞系的建立和表征”J Ext Clin Cancer Res.. 19(1)。

Yamaguchi K.et al.: "Inhibitory effect of TNP-470 on lymph node metastasis of human gastric carcinoma line established by orthotopic implantation."Tumor Res.. 36. 15-21 (2001)

Yamaguchi K.et al.：“TNP-470 对原位植入建立的人胃癌系淋巴结转移的抑制作用。”Tumor Res.. 36. 15-21 (2001)

Nishimori H, Yasoshima T, Denno R, Shishido T, Hata F, Okada Y, Ura H, Yamaguchi K, Isomura H, Sato N and Hirata K: "A novel experimental mouse model of peritoneal dissenmination of human gastric cancer cells; Different mechanism in peritoneal disseminati

Nishimori H、Yasoshima T、Denno R、Shishido T、Hata F、Okada Y、Ura H、Yamaguchi K、Isomura H、Sato N 和 Hirata K：“人胃癌细胞腹膜传播的新型实验小鼠模型；不同的机制

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