Analysis of micro and molecular mechanism of lymphatic metastasis, and development of new strategy targeting for tumor lymphatic vessel

淋巴转移微观分子机制分析及肿瘤淋巴管靶向新策略开发

• 批准号: 15390370
• 负责人: MIZOI Takayuki
• 金额: $ 9.15万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390370/
• 关键词: colorectal cancer; lymph node metastasis; lymphangiogenesis; CD44; hyarulonic acid; cell attachment; microarray

We investigated the interaction between CD44 and its ligand, hyaluronic acid, on lymph node metastasis of colorectal cancers. In 90% of nude mice, lymph node metastasis was established by alloic transplantation of LS174T and LS174T-NEO, which expressed high levels of CD44. In contrast, Colo320 and LS174T-AS2, which were transfected with antisense CD44 gene, had no expressions of CD44 and never metastasized to lymph nodes by alloic transplantation in nude mice. In immunohistochemistry of lymphatic vessels, LS174T-NEO showed marked lymphatic invasion, in contrast to minimal lymphatic invasion in LS174T-AS2 and Colo320. LS174T and LS174T-NEO widely attached to normal lymph nodes, and it was inhibited by anti-hyaluronic acid processing. On the other hand, the attachment to lymph nodes was significantly reduced in LS174T-AS2.We evaluated the ability of colorectal cancer cell lines on metastasis to lymph nodes, and metastatic (or anti-metastatic) genes were screened by cDNA microarray. Twenty three of human colorectal cancer cell lines were investigated on the lymph node metastasis in alloic transplantation model and the incidence of metastasis was varied widely as follows : KM12c 100%(7/7), CloneA 100%(9/9), HT29 94.4%(17/18),..., SW1116 25%(2/8), DLD1 0%(0/13). Cell lines were divided into three groups (high, middle and low) according to the incidence of lymph node metastasis. Based on cDNA microarray analysis together with in vivo experiments, 107 candidates of metastasis-related genes were extracted. The expression levels of some of those genes were confirmed elevated or depressed with real-time RT-PCR as well. Further, there was a relationship between expression levels of candidate genes and clinical lymph node metastasis in colorectal cancers.

我们研究了 CD44 与其配体透明质酸之间的相互作用对结直肠癌淋巴结转移的影响。在90%的裸鼠中，通过表达高水平CD44的LS174T和LS174T-NEO的同种异体移植建立了淋巴结转移。相反，转染反义CD44基因的Colo320和LS174T-AS2不表达CD44，并且在裸鼠同种异体移植中从未发生淋巴结转移。在淋巴管的免疫组织化学中，LS174T-NEO 显示出明显的淋巴浸润，​​而 LS174T-AS2 和 Colo320 中的淋巴浸润极少。 LS174T和LS174T-NEO广泛附着于正常淋巴结，并受到抗透明质酸加工的抑制。另一方面，LS174T-AS2对淋巴结的附着显着减少。我们评估了结直肠癌细胞系向淋巴结转移的能力，并通过cDNA微阵列筛选转移（或抗转移）基因。研究了23种人结直肠癌细胞系同种异体移植模型的淋巴结转移情况，其转移率差异较大：KM12c 100%(7/7)、CloneA 100%(9/9)、HT29 94.4%(17/18)、...、SW1116 25%(2/8)、DLD1 0%(0/13)。根据淋巴结转移发生率将细胞系分为高、中、低三组。基于cDNA微阵列分析和体内实验，提取了107个候选转移相关基因。实时 RT-PCR 也证实其中一些基因的表达水平升高或降低。此外，候选基因的表达水平与结直肠癌的临床淋巴结转移之间存在关系。

项目成果

Characterization of the organic cation transporter SLC22A16: A doxorubicin importer

• DOI: 10.1016/j.bbrc.2005.05.174
• 发表时间: 2005-08-05
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Okabe, M;Unno, M;Abe, T
• 通讯作者: Abe, T

Intraepithelial CD8+ T-cell-count becomes a prognostic factor after a longer follow-up period in human colorectal carcinoma: possible association with suppression of micrometastasis.

• DOI: 10.1038/sj.bjc.6602201
• 发表时间: 2004-11-01
• 期刊: BRITISH JOURNAL OF CANCER
• 影响因子: 8.8
• 作者: Chiba, T;Ohtani, H;Mizoi, T;Naito, Y;Sato, E;Nagura, H;Ohuchi, A;Ohuchi, K;Shiiba, K;Kurokawa, Y;Satomi, S
• 通讯作者: Satomi, S

Thymidine phosphorylase expressed in macrophages enhances antitumor effect of 5'-deoxy-5-fluorouridine on human colorectal carcinoma cells.

巨噬细胞中表达的胸苷磷酸化酶增强 5-脱氧-5-氟尿苷对人结直肠癌细胞的抗肿瘤作用。

• 发表时间: 2003
• 期刊: Anticancer Res 23
• 作者: Zhang J;et al.
• 通讯作者: et al.

Thymidine phosphorylase expressed in macrophages enhances effect of 5'-deoxy-5-fluorouridine on humancolorectal carcinoma cells.

巨噬细胞中表达的胸苷磷酸化酶增强 5-脱氧-5-氟尿苷对人结直肠癌细胞的作用。

• 发表时间: 2003
• 期刊: Anticancer Res 23
• 作者: Zhang J;et al.
• 通讯作者: et al.