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The role of E1AF, an ets-oncogene family tranascription factor in cell cycle regulation.

E1AF（一种 ets 癌基因家族转录因子）在细胞周期调节中的作用。

基本信息

批准号：
12470431
负责人：
TOTSUKA Yasunori
金额：
$ 9.6万
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：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470431/
关键词：
E1AF p21 p53 cell cycle regulation 細胞周期調節

项目摘要

E1AF is a member of the ets-oncogene family transcription factor, and E1AF was shown to bind to the promoter region of MMPs that induce transcription from multiple MMP genes. We demonstrated that Ad type 12 E1A is able to suppress the invasion ability of a cancer cell line by inactivating E1AF expression. Ad12E1A expressing oral squamous cell carcinoma cell line was established. Northern blotting indicated that the expression of E1AF mRNA decreased in transfected cells. Reduced expression of MMPs was identified both in mRNA levels and in protein levels. Reduced transcriptional activity of MMP gene was confirmed by CAT assay. These results suggest that Ad12E1A downregulates E1AF which, in turn, restrains invasion ability of human cancer cells by way of downregulaton of MMP expression.Ca9.22 cells that manifest low levels of E1AF and MMP-1 and -9 expression were transfected with metallothionein inducible E1AF expression vector. MMP-9 and p21_<waf1/cipl> protein were synergistically increased in ZnCl stimulated MT9.22 cells, and MMP-9 and p21 protein were expressed in the same cells when E1AF was induced in MT9.22 cells.These results imply that invasive growth of tumor cells occur in static state of cell cycle and E1AF play key role of this phenomenon.Luciferase assays, using different amounts of E1AF and p53 expression vectors along with p2 1 promoter luciferase reporter plasmids, demonstrate an increase in the activity of p21. Colony formation assay indicates that E1AF is capable of tumor suppression similar to p53. These results, therefore, suggest that E1AF is able to interact synergistically with p53 ; in consequence, this interaction can influence p21 leading to an enhancement of its activity and eventually, tumor suppression.

E1AF是ets-癌基因家族转录因子的一员，E1AF被证明与MMPs的启动子区域结合，诱导多个MMP基因的转录。我们证明了Ad 12型E1A能够通过使E1AF表达失活来抑制癌细胞系的侵袭能力。建立表达Ad12E1A的口腔鳞状细胞癌细胞系。Northern blotting结果显示，转染细胞中E1AF mRNA的表达降低。在mRNA水平和蛋白水平上均发现MMPs的表达降低。CAT实验证实MMP基因转录活性降低。这些结果表明，Ad12E1A下调E1AF，进而通过下调MMP表达抑制人癌细胞的侵袭能力。ca9.22细胞表达E1AF和MMP-1、-9低水平，转染金属巯基蛋白诱导的E1AF表达载体。MMP-9和p21_<waf1/cipl>蛋白在ZnCl刺激的MT9.22细胞中协同升高，而在E1AF诱导的MT9.22细胞中，MMP-9和p21蛋白在相同的细胞中表达。这些结果表明肿瘤细胞的侵袭性生长发生在细胞周期的静态状态，而E1AF在这一现象中起着关键作用。使用不同数量的E1AF和p53表达载体以及p21启动子荧光素酶报告质粒进行荧光素酶检测，显示p21的活性增加。集落形成实验表明E1AF具有类似于p53的抑瘤作用。因此，这些结果表明E1AF能够与p53协同作用；因此，这种相互作用可以影响p21，从而增强其活性并最终抑制肿瘤。