ヒト・ホメオドメインタンパクによる新しいカドヘリン遺伝子の発現制御機構の解析

人同源域蛋白新的钙粘蛋白基因表达调控机制分析

• 批准号: 06670240
• 负责人: TANIGUCHI Yasushi
• 金额: $ 1.41万
• 依托单位: Tokai University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670240/
• 关键词: homeobox; HOXD3; HEL cells; transfectants; expression vectors; integrin beta3; R-cadherin; cell adhesion; インテグリンαIIbβ3

We isolated the human homeobox gene HOXD3 on chromosome 2 from a human genomic library and determined its nucleotide sequence. In the present study, in order to study the role of the HOXD3 gene in erythropoiesis, expression vectors containing the HOXD3 gene inthe sense or antisense orientation were introduced into HEL cells. The sensetransfectants overexpressing the HOXD3 gene formed aggregates, which were composed of densely associated cells adhering to tissue-culture dishes, whereas the parental HEL cells and antisense transfectants adhered poorly to the dishes. Furthermore, the sense transfectants overexpressing the HOXD3 gene attached more efficiently to fibronectin and collagen than did the antisense transfectants and parental HEL cells. Northern blot analysis showed that integrin beta3 mRNA levels were significantly increased in the HEL cells overexpressing the HOX4A gene. FACS analysis showed that the sense transfectants expressed increased levelsof integrin alpha II bbeta3 complex. In addition, the antisense transfectants expressed R-cadherin more abundantly than the sense transfectants and parental HEL cells. These results implicate the homeobox gene HOXD3 in the regulation of cell abhesion processes.

我们从人类基因组文库中分离了人类2号染色体上的同源异型盒基因HOXD 3，并测定了其核苷酸序列。为了研究HOXD 3基因在红细胞生成中的作用，本研究将HOXD 3基因的正、反义表达载体导入HEL细胞。过表达HOXD 3基因的正义转染子形成聚集体，其由粘附于组织培养皿的密集相关细胞组成，而亲本HEL细胞和反义转染子粘附于皿较差。此外，过表达HOXD 3基因的正义转染子比反义转染子和亲本HEL细胞更有效地附着于纤连蛋白和胶原。北方印迹分析显示，整合素β 3的mRNA水平显着增加HEL细胞中过表达HOX 4A基因。流式细胞仪分析显示正义转染子表达的整合素α Ⅱ b β 3复合物水平增加。此外，反义转染细胞表达的R-钙粘蛋白比正义转染细胞和亲本HEL细胞更丰富。这些结果暗示同源异型盒基因HOXD 3在调节细胞粘附过程。

项目成果

Taniguchi, Y., Katsumata, Y., Koido, S., Suemizu, H., Yoshimura, S., Moriuchi, T., Okumura, K., Kagotani, K., Taguchi, H., imanishi, T., Gojobori, T., and Inoko, H.: "Cloning, sequencing, and chromosomal localization of two tandemly arranged human pseudog

谷口，Y.，胜又，Y.，小户，S.，苏水，H.，吉村，S.，森内，T.，奥村，K.，鹿谷，K.，田口，H.，今西，T.，

Taniguchi, Y., Komatsu, N., andMoriuchi, T.: "Overexpression of the HOX4A (HOXD3) homeobox gene in human erythroleukemia HEL cells results in altered adhesive properties" Blood. 85. 2786-2794 (1995)

Taniguchi, Y.、Komatsu, N. 和 Moriuchi, T.：“人红白血病 HEL 细胞中 HOX4A (HOXD3) 同源框基因的过度表达导致粘附特性改变”血液。

Taniguchi,Y.: "Overexpression of the HOX4A(HOXD3) homeobox gene in human erythroleukemia HEL cells results in altered adhesive properties." Blood,in press.

Taniguchi, Y.：“人红白血病 HEL 细胞中 HOX4A (HOXD3) 同源盒基因的过度表达会导致粘附特性改变。”

Arimori K.: "Nucleotide sequence of a human genomic DNA fragment containing the PCNA pseudogene and its localization on chromosome 4." Tokai Exp.Clin.Med.19. 1-5 (1994)

Arimori K.：“含有 PCNA 假基因的人类基因组 DNA 片段的核苷酸序列及其在 4 号染色体上的定位。”

Arimori, K.: "Nucleotide sequence of a human genomic DNA fragment containing the PCNA pseudogene and its localization on chromosome 4." Tokai Exp. Clin. Med.19. 1-5 (1994)

Arimori, K.：“含有 PCNA 假基因的人类基因组 DNA 片段的核苷酸序列及其在 4 号染色体上的定位。”