Cyclin D1 inhibits cell proliferation through binding to PCNA and Cdk2
Cyclin D1 inhibits cell proliferation through binding to PCNA and Cdk2
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DOI：
10.1006/excr.1998.4306
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发表时间：
1999-02-01
影响因子：
3.7
通讯作者：
Mitsui, Y
中科院分区：
文献类型：
--
作者：
Fukami-Kobayashi, J;Mitsui, Y
Cyclin D1 被认为是细胞生长的促进因子。然而，我们之前表明，体外衰老人成纤维细胞中细胞周期蛋白 D1 的表达显着增加。在这里，我们研究 cyclin D1 的过度表达是否会抑制细胞增殖。与使用模拟和细胞周期蛋白 E 表达载体相比，在 NIH-3T3、TIG-1、CHO-K1 和 HeLa 细胞中，转染细胞周期蛋白 D1 表达载体后的集落形成受到抑制。瞬时转染实验表明，cyclin D1 的过度表达会抑制 TIG-1 细胞的 DNA 合成。与年轻细胞相比，衰老细胞中细胞周期蛋白 D1 与 PCNA 和 cdk2 的复合物显着增加。体外，过量的谷胱甘肽 S-转移酶 (GST)-细胞周期蛋白 D1 会抑制 DNA 复制并抑制 cdk2 依赖性激酶活性。带有 PCNA 或 cdk2 表达载体的 NIH-3T3 转染子的 DNA 合成不受细胞周期蛋白 D1 过表达的抑制。这些结果表明，过量的细胞周期蛋白 D1 通过与 PCNA 和 cdk2 结合来抑制 DNA 复制和 cdk2 活性，从而抑制细胞增殖，就像在衰老细胞中一样。 (C) 1999 年学术出版社。
Cyclin D1 is known as a promoting factor for cell growth. We previously showed, however, that the expression of cyclin D1 increases markedly in senescent human fibroblasts in vitro. Here we investigate whether the overexpression of cyclin D1 inhibits cell proliferation. Colony formation after transfection with the cyclin D1 expression vector was repressed in NIH-3T3, TIG-1, CHO-K1, and HeLa cells, compared with those with mock and cyclin E expression vectors. A transient transfection assay demonstrated that the overexpression of cyclin D1 inhibited DNA synthesis of TIG-1 cells. The complexes of cyclin D1 with PCNA and cdk2 increased remarkably in senescent cells, compared with young counterparts. Excessive glutathione S-transferase (GST)-cyclin D1 inhibited DNA replication and repressed cdk2-dependent kinase activity in vitro. DNA synthesis of NIH-3T3 transfectants with PCNA or cdk2 expression vectors was not inhibited by the overexpression of cyclin D1. These results indicate that an excessive level of cyclin D1 represses cell proliferation by inhibiting DNA replication and cdk2 activity through the binding of cyclin D1 to PCNA and cdk2, as it does in senescent cells. (C) 1999 Academic Press.