Function of novel gene, EAT that is early gene induced by all-trans retinoic acid in human EC cells

全反式维甲酸诱导的早期基因EAT在人EC细胞中的功能

• 批准号: 08457066
• 负责人: HATA Junichi
• 金额: $ 4.86万
• 依托单位: KEIO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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-08457066/
• 关键词: Human EC cells; EAT; mcl1; Apoptosis; BCL-2; 10(1) cell; Transgenic mice; Germ cell tumors; CDDP; モノクローナル抗体; CDDP

The aim of this study was to isolate the essential genes in retinoic acid (RA)-induced differentiation of human EC cells, NCR-G3 cell. We isolated a novel gene, EAT which transiently expressed in the early stage of differentiation and has homology with bcl-2, a cell death-preventing molecule. We obtained following results by analyzing this novel gene, EAT.To elucidate expression of EAT in normal and fetal tissue, a murine monoclonal antibody, 3A2 was generated by use of recombinant protein to EAT. By immnohistochemical studies, EAT expression was observed in a wide variety of human fetal and adult tissues, including those of epithelial, hematopoietic and endocrine cells. In murine embryos, EAT was induced rapidly after fertilization reaching maximum level at the from the 2 - to 8-cell stage and then decrease to below unfertilized egg levels in blastocyst.2) We determined whether the apoptotic cell death induced by chemotherapeutic agents could be inhibited by EAT as is reported with Bcl-2. Cells transfected with EAT showed higher resistance to CDDP or among other chemotherapeutic agents tested. DNA fragmentation of the parental cells following treatment with CDDP and carboplatin was observed in a concentration-dependent manner. In contrast, cells transfected with EAT did not show DNA fragmentation following treatment with the same concentration of these drugs. Acute tubular necrosis induced by CDDP administration did not occur in the EAT transgenic mice.These results suggest that EAT may play an important role in early embryogenesis and sequential cell function by its anti-apoptotic functions.

本研究的目的是分离维甲酸（RA）诱导人食管癌细胞NCR-G3分化的关键基因。我们分离了一个新基因EAT，它在分化早期瞬时表达，与细胞死亡预防分子bcl-2具有同源性。为了阐明EAT在正常组织和胎儿组织中的表达，我们利用重组蛋白制备了抗EAT的鼠单克隆抗体3A 2。通过免疫组化研究，EAT在多种人类胎儿和成人组织中表达，包括上皮细胞、造血细胞和内分泌细胞。在小鼠胚胎中，EAT在受精后迅速诱导，在2 -至8-细胞期达到最大水平，然后在囊胚中降低至低于未受精卵水平。2）我们确定了由化疗剂诱导的凋亡性细胞死亡是否可以被EAT抑制，如Bcl-2所报道的。用EAT转染的细胞显示出对CDDP或其他测试的化疗剂的更高抗性。用CDDP和卡铂处理后，以浓度依赖性方式观察到亲本细胞的DNA片段化。相反，用EAT转染的细胞在用相同浓度的这些药物处理后没有显示DNA片段化。EAT转基因小鼠未出现CDDP诱导的急性肾小管坏死，提示EAT可能通过其抗凋亡功能在早期胚胎发生和细胞功能中发挥重要作用。

项目成果

Ogata,T.: "Impaired male sex development in an infnat with molecularly defined partial 9p monosomy: implication for a testis forming gene(s) on 9P"J Med Genet. 34. 331-334 (1997)

Ogata,T.：“具有分子定义的部分 9p 单体的 infnat 中男性性发育受损：对 9P 上睾丸形成基因的影响”J Med Genet。

Okita, H., Umezawa, A., Hata, J.: "Up-regulated expression of murine Mcl1/EAT, aBcl-2 related gene, in the early stage of differentiation of murine embryonal carcinoma cells and embryonic stem cells."BBA. 1398. 335-341 (1998)

Okita, H.、Umezawa, A.、Hata, J.：“在小鼠胚胎癌细胞和胚胎干细胞分化的早期阶段，小鼠 Mcl1/EAT、aBcl-2 相关基因的表达上调。”BBA

Maruyma,T.: "Heat shock induces differentiation of human embryonal cacinoma cells into trophoectodermal lineages." Exp.Cell Res.224 :. 123-127 (1996)

Maruyma,T.：“热休克诱导人胚胎癌细胞分化为滋养外胚层谱系。”

Sugimono,T: "Neurogenic potential of Ewing's sarcoma cells"Virchow Archiv. 430. 41-46 (1997)

Sugimono，T：“尤文氏肉瘤细胞的神经源潜力”Virchow Archiv。

Maruyama, T., Umezawa, A., Hata, J.: "heat shock induces differentiation of human embryonal carcinoma cells into trophoectodermal lineages."Exp. Cell Res.. 224. 123-127 (1996)

Maruyama, T.、Umezawa, A.、Hata, J.：“热休克诱导人胚胎癌细胞分化为滋养外胚层谱系。”Exp。