一种锌指蛋白的特性及其与前列腺癌细胞凋亡的关联

Characterization of a zinc-finger protein and its association with apoptosis in prostate cancer cells.

作者信息

Chang G T, Steenbeek M, Schippers E, Blok L J, van Weerden W M, van Alewijk D C, Eussen B H, van Steenbrugge G J, Brinkmann A O

机构信息

Department of Endocrinology and Reproduction, Josephine Nefkens Institute, Erasmus University Rotterdam, The Netherlands.

出版信息

J Natl Cancer Inst. 2000 Sep 6;92(17):1414-21. doi: 10.1093/jnci/92.17.1414.

DOI:10.1093/jnci/92.17.1414

PMID:10974077

Abstract

BACKGROUND

The transition from androgen-dependent to androgen-independent prostate cancer is not fully understood but appears to involve multiple genetic changes. We have identified a gene, GC79, that is more highly expressed in androgen-dependent LNCaP-FGC human prostate cancer cells than in androgen-independent LNCaP-LNO human prostate cancer cells. Physiologic levels (0.1 nM:) of androgens repress expression of GC79 messenger RNA (mRNA) in LNCaP-FGC cells. To determine the role of GC79, we cloned its complementary DNA (cDNA) and functionally characterized its product.

METHODS

The differentially expressed GC79 gene was cloned from human prostate cDNA libraries, sequenced, and transfected into mammalian cells to study its function. Expression of GC79 was analyzed in various adult and fetal human tissues and in prostate glands of castrated rats. The association of GC79 expression and apoptosis was investigated in COS-1 and LNCaP cells transfected with GC79 cDNA. All statistical tests are two-sided.

RESULTS

Sequence analysis indicates that GC79 encodes a large, complex, multitype zinc-finger protein, containing nine C(2)H(2)-type zinc-finger domains, a cysteine-rich region, and a GATA C(4)-type zinc-finger domain. Castration-induced androgen withdrawal increased the expression of GC79 mRNA in the regressing rat ventral prostate, suggesting that the expression of GC79 mRNA is associated with the process of apoptotic cell death in the rat ventral prostate. Transfection and induction of GC79 cDNA in both COS-1 and LNCaP prostate cancer cells led to an apoptotic index that was eightfold higher (P:<.001, two-sided Student's t test) than that observed in uninduced transfected cells.

CONCLUSIONS

We have cloned an androgen-repressible gene, GC79, that is potentially involved in apoptosis. This finding may have implications for the development of androgen-independent prostate cancer and, ultimately, for the treatment of prostate cancer.

摘要

背景

从雄激素依赖型前列腺癌向雄激素非依赖型前列腺癌的转变尚未完全明确，但似乎涉及多种基因变化。我们鉴定出一个基因GC79，其在雄激素依赖型LNCaP-FGC人前列腺癌细胞中的表达高于雄激素非依赖型LNCaP-LNO人前列腺癌细胞。雄激素的生理水平（0.1 nM）可抑制LNCaP-FGC细胞中GC79信使核糖核酸（mRNA）的表达。为确定GC79的作用，我们克隆了其互补脱氧核糖核酸（cDNA）并对其产物进行了功能表征。

方法

从人前列腺cDNA文库中克隆差异表达的GC79基因，进行测序，并转染至哺乳动物细胞以研究其功能。分析了GC79在各种成人和胎儿人体组织以及去势大鼠前列腺中的表达。在转染了GC79 cDNA的COS-1和LNCaP细胞中研究了GC79表达与细胞凋亡的关联。所有统计检验均为双侧检验。

结果

序列分析表明，GC79编码一种大型、复杂的多类型锌指蛋白，包含9个C2H2型锌指结构域、一个富含半胱氨酸的区域和一个GATA C4型锌指结构域。去势诱导的雄激素撤退增加了退行性大鼠腹侧前列腺中GC79 mRNA的表达，表明GC79 mRNA的表达与大鼠腹侧前列腺中的凋亡细胞死亡过程相关。在COS-1和LNCaP前列腺癌细胞中转染并诱导GC79 cDNA导致凋亡指数比未诱导的转染细胞高8倍（P<0.001，双侧Student t检验）。