The destiny of the neural crest cells where the homeobox gene was ectopically expressed.

同源盒基因异位表达的神经嵴细胞的命运。

• 批准号: 08670036
• 负责人: TANIGUCHI Yasushi
• 金额: $ 1.66万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670036/
• 关键词: homeobox; HOXD3; cell adhesion; integrin; cadherin; Wnt-1; neural crest; transgenic mouse; トランスジェニックマウス; インテグリンβ3; R-カドヘリン; トアンスジェニックマウス; 細胞接着因子

I isolated the human homeobox gene HOXD3 on chromosome 2 from a human genomic library and determined its nucleotide sequence. To study the role of HOXD3 gene in erythropoiesis, expression vectors containing the HOXD3 gene in the sense or antisense orientation were introduced into HEL cells. 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 HOXD3 gene, whereas the integrin beta1 and alphaIIb mRNA levels did not show a distinct correlation with HOXD3 mRNA levels. Fluorescense-activated cell sorting analysis showed that the sense transfectants overexpressing the HOXD3 gene expressed increased levels of integrin alphaIIbbeta3 (GP IIbIIIa) complex as compared with the parental HEL cells and antisense transfectants. In addition, reverse transcriptase-mediated polymerase chain reaction (RT-PCR) analysis revealed that R-cadherin mRNA Ievels were significantly increased in the antisense transfectants expressing very low levels of the HOXD3 mRNA as compared to the sense transfectants and parental HEL cells. These results implicate the homeobox gene HOXD3 in the regulation of cell adhesion process. To verify the regulation by the HOXD3 also in developing mouse embryos, I made transgenic mice where the human HOXD3 gene was specifically expressed in the neural crest cells and analyzed the differentiation and adhessiveness of the cells.

我从人类基因组文库中分离了2号染色体上的人类同源异型盒基因Hoxd3，并测定了它的核苷酸序列。为了研究Hoxd3基因在红细胞生成中的作用，将Hoxd3基因正、反义定向的表达载体导入HEL细胞。高表达Hoxd3基因的正义转染体与纤维连接蛋白和胶原的结合效率高于反义转染组和亲本HEL细胞。Northern印迹分析显示，在Hoxd3基因过表达的HEL细胞中，整合素Beta3mRNA水平显著升高，而整合素Beta1和AlphaIIb mRNA水平与Hoxd3mRNA水平无明显相关性。荧光激活的细胞分类分析表明，与亲本HEL细胞和反义转染体相比，高表达Hoxd3基因的正义转染体表达的整合素αIIbIIIa(GP IIbIIIa)复合体水平增加。此外，逆转录酶介导的聚合酶链式反应(RT-PCR)分析显示，与正义转染体和亲本HEL细胞相比，表达极低水平Hoxd3的反义转染体的R-钙粘素mRNA水平显著升高。这些结果表明同源异型盒基因Hoxd3参与了细胞黏附过程的调控。为了验证Hoxd3基因在小鼠胚胎发育中的调节作用，我制作了人类Hoxd3基因在神经脊细胞中特异表达的转基因小鼠，并分析了细胞的分化和粘附性。

项目成果

Taniguchi, Y.: "Regulation of the expression of cell adhesion molecules by the homeobox gene HOXD3 in human leukemia cells." The cell. 28. 250-254 (1996)

Taniguchi, Y.：“人类白血病细胞中同源框基因 HOXD3 对细胞粘附分子表达的调节。”

Taniguchi, Y.: "Cloning,sequencing,and chromosomal localization of two tandemly arranged human pseudoganes for the proliferating cell nuclear antigeu (RCNA)" Manualian Genome. 7. 906-908 (1996)

Taniguchi, Y.：“增殖细胞核抗原 (RCNA) 的两个串联排列的人类假细胞的克隆、测序和染色体定位”手册基因组。

Taniguchi, Y.: "Nucleotide sequence of the Ring3 gene in the class II region of the mouse MHC and its abundant expression in testicular germ cells." Genomics. 印刷中. (1998)

Taniguchi, Y.：“小鼠 MHC II 类区域的 Ring3 基因的核苷酸序列及其在睾丸生殖细胞中的丰富表达”（正在出版）。

谷口 泰史: "ホメオボックス遺伝子HOXD3によるヒト白血病細胞接着分子発現の制御" 細胞. 28. 250-254 (1996)

Yasushi Taniguchi：“同源框基因 HOXD3 对人类白血病细胞粘附分子表达的调节”Cell。28. 250-254 (1996)

Y.Taniguchi: "Cloning,sequencing,and chromosomal localization of two tandenly arranged human psedogenes for the preliferating cell nuclear antigen" Mammalian Genome. 7/12. 906-908 (1996)

Y.Taniguchi：“增殖细胞核抗原的两个串联排列的人类假基因的克隆、测序和染色体定位”哺乳动物基因组。