Analysis of Master Genes Associated with Invasion and Metastasis by Malignant Melanoma

恶性黑色素瘤侵袭和转移相关主基因分析

• 批准号: 13470377
• 负责人: YAMAMOTO Yuhei
• 金额: $ 9.02万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470377/
• 关键词: malignant melanoma; homeobox genes; antisense; invasion; flow cytometry; cDNA microarray

Homeobox genes regulate sets of genes that determine cellular fates in embryonic morphogenesis and maintenance of adult tissue architecture by regulating cellular motility and cell-cell interactions. Our previous studies showed that a specific member, HOXD3, when overexpressed, enhanced cell motility and invasiveness of human lung cancer A549 cells (Hamada et al. Int. J. Cancer 2001; 93: 516-25 [19]). In the present study, we investigated the roles of HOXD3 in motile and invasive behavior of human malignant melanoma cells. Of seven melanoma cell lines examined here, six cell lines expressed the HOXD3 gene, whereas normal melanocytes did not. We transduced the HOXD3-antisense gene expression vector into two cell lines (A375M and MMIV). The cell transduced with the HOXD3-antisense gene showed reduced in vitro invasion of Matrigel. The transduction of the HOXD3-antisense gene also decreased cell spreading, haptotactic activity to vitronectin and laminin-1, and phagokinetic activity. To find the difference of gene expression between the HOXD3-antisense-transduced A375M cells and the control A375MNeo2 cells, we carried out cDNA microarray analysis. The results of the microarray analysis indicated that the increased expression of cdc42-interacting protein 4, KIAA0554 and tropomyosin 1, which are all associated with the cytoskeletal system, may be involved in the reduction of motile and invasive activity by the HOXD3-antisense gene transduction.We hypothesized that HOX genes give the cells spatial information to maintain tissue-or organ-specificity in adult body and that the deregulated expression of HOX genes results in tumor development. To better understand the roles of HOX genes in melanoma genesis and subsequent malignant progression, we compared the HOX gene expression patterns among melanomas in different body sites.

同源框基因通过调节细胞运动和细胞间的相互作用来调节一组基因，这些基因决定了胚胎形态发生和成人组织结构的维持中的细胞命运。我们以前的研究表明，当一个特定的成员Hoxd3过表达时，可以增强人肺癌A549细胞的细胞运动性和侵袭力(Hamada等人。内部J.癌症2001；93：516-25[19])。在本研究中，我们研究了Hoxd3在人恶性黑色素瘤细胞运动和侵袭行为中的作用。在这里研究的七个黑色素瘤细胞系中，有六个细胞系表达Hoxd3基因，而正常的黑素细胞不表达。我们将Hoxd3反义基因表达载体导入A375M和MMIV细胞中。转导Hoxd3反义基因的细胞体外侵袭Matrigel的能力降低。Hoxd3反义基因的转导也降低了细胞铺展、对Vitronectin和Laminin-1的趋化活性以及吞噬活性。为了找出Hoxd3反义基因转导的A375M细胞和对照A375MNeo2细胞基因表达的差异，我们进行了基因芯片分析。芯片分析结果表明，与细胞骨架系统相关的CDC42相互作用蛋白4、KIAA0554和原肌球蛋白1的表达增加可能与HOXD3反义基因转导降低运动和侵袭活性有关。我们假设HOX基因为细胞提供空间信息，以维持成人体内的组织或器官特异性，并且HOX基因的表达失控导致肿瘤的发生。为了更好地了解HOX基因在黑色素瘤发生和随后的恶性进展中的作用，我们比较了不同部位黑色素瘤中HOX基因的表达模式。

项目成果

大久保 佳子: "ホメオボックス遺伝子,HOXD3アンチセンス発現ベクター導入によるヒト悪性黒色腫細胞のinvitro浸潤性の低下"北海道医学雑誌. 76巻4号. 239-250 (2001)

Yoshiko Okubo：“通过引入同源框基因、HOXD3反义表达载体减少人恶性黑色素瘤细胞的体外侵袭性”北海道医学杂志，第76卷，第4期。239-250（2001）

Okubo Yoshiko: "Transduction of HOXD3-antisense into human melanoma cells results in decreased invasive and motile activities."Clinical & experimental metastasis. 19(6). 503-511 (2002)

Okubo Yoshiko：“将 HOXD3 反义转导到人类黑色素瘤细胞中会导致侵袭性和运动性活动减少。” 临床

Yoshiko Okubo, Jun-ichi Hamada, Yoko Takahashi, Mitsuhiro Tada, Arata Tsutsumida, Keiji Furuuchi, Tetsuya Aoyama, Tsuneki Sugihara, Tetsuya Moriuchi: "Transduction of HOXD3-antisense into human melanoma cell results in decreased invasive and activities."C

Yoshiko Okubo、Jun-ichi Hamada、Yoko Takahashi、Mitsuhiro Tada、Arata Tsutsumida、Keiji Furuuchi、Tetsuya Aoyama、Tsuneki Sugihara、Tetsuya Moriuchi：“将 HOXD3 反义转导到人类黑色素瘤细胞中会导致侵袭性和活性降低。”C

Okubo Yoshiko: "Transduction of HOXD3-antisense into human melanoma cells results in decreased invasive and motile activities"Clinical & experimental metastasis. 19(6). 503-511 (2002)

Okubo Yoshiko：“将 HOXD3 反义转导到人黑色素瘤细胞中会导致侵袭性和运动性活动减少”临床

大久保 佳子: "ホメオボックス遺伝子 HOXD3 アンチセンス発現ベクター導入によるヒト悪性黒色腫細胞の in vitro 浸潤性の低下"Skin Cancer. 76巻4号. 239-250 (2001)

Yoshiko Okubo：“通过引入同源盒基因 HOXD3 反义表达载体减少人类恶性黑色素瘤细胞的体外侵袭性”，皮肤癌，第 76 卷，第 4 期，239-250（2001 年）。