Inhibition of cell proliferation by the interferon-inducible 204 gene, a member of the Ifi 200 cluster
Inhibition of cell proliferation by the interferon-inducible 204 gene, a member of the Ifi 200 cluster
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DOI：
10.1038/sj.onc.1201677
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发表时间：
1998-03-26
期刊：
影响因子：
8
通讯作者：
Landolfo, S
中科院分区：
文献类型：
--
作者：
Lembo, M;Sacchi, C;Landolfo, S
通过将组成型表达p204的表达载体转染到几种细胞系中，研究了IFN-诱导型p204作为生长调节剂的作用。如细胞灶试验所示，与PRE和p107一样，p204在敏感细胞中是有效的生长抑制剂。由于组成型过表达p204的敏感细胞系的稳定转染子不能在体外建立，我们将204 cDNA插入到带有重金属诱导型启动子的载体中，在此我们表明，缺乏内源性p204的B6 MEF成纤维细胞的增殖被细胞核中的瞬时p204表达强烈抑制，p204延迟G（1）进展到S期，并且细胞积累的DNA含量相当于停滞在G（1）晚期的细胞。此外，通过产生其中Ifi 204基因在所有组织中组成型表达的转基因小鼠来研究p204在体内细胞生长控制中的作用。为此，构建了在SV 40病毒启动子控制下的携带204 cDNA的表达载体，通过将这些载体注射到受精小鼠卵中实现p204转基因的过表达，在体外随访中与胚胎发育至四细胞期相容。然而，没有获得具有转基因的完整拷贝的存活动物，这表明高水平和组成型水平的p204表达可以损害正常胚胎发育。这些发现表明p204在生长调节中起负作用，并提供了关于IFN抑制细胞增殖的分子机制的新信息。
The role of the IFN-inducible p204 as growth regulator was investigated by transfecting an expression vector constitutively expressing p204 into several cell lines, Like PRE and p107, p204 is a potent growth inhibitor in sensitive cells, as demonstrated by the cell focus assay, Since stable transfectants of sensitive lines constitutively overexpressing p204 could not be established in vitro, we inserted the 204 cDNA into a vector bearing an heavy-metal-inducible promoter, Here we show that proliferation of B6MEF fibroblasts lacking endogenous p204 is strongly inhibited by transient p204 expression in the nucleus, p204 delays G(1) progression into the S-phase and cells accumulate with a DNA content equivalent to cells arrested in late G(1). Moreover, the role of p204 in the control of cell growth in vivo was investigated by generating transgenic mice in which the Ifi 204 gene was constitutively expressed in all tissues, To this end, expression vectors bearing the 204 cDNA under the control of the SV40 viral promoter were constructed, The overexpression of the p204 transgene achieved by injecting fertilized mouse eggs with these vectors was compatible with embryo development up to the four-cell stage in an in vitro follow-up of 4.5 days, However, no viable animals with an intact copy of the transgene were obtained, suggesting that high and constitutive levels of p204 expression can impair normal embryo development, These findings indicate that p204 plays a negative role in growth regulation and provide new information about the molecular mechanisms exploited by IFNs to inhibit cell proliferation.