Androgen-induced proliferative quiescence in prostate cancer cells: The role of AS3 as its mediator
Androgen-induced proliferative quiescence in prostate cancer cells: The role of AS3 as its mediator
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DOI：
10.1073/pnas.97.18.10185
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发表时间：
2000-08-29
影响因子：
11.1
通讯作者：
Soto, AM
中科院分区：
文献类型：
--
作者：
Geck, P;Maffini, MV;Soto, AM
在成年哺乳动物的前列腺中，大多数上皮细胞处于增殖停滞状态。雄激素通过诱导细胞周期停滞于G(0)/G(1)期来调节这一效应。通过消减的cDNA文库鉴定了雄激素诱导的这种增殖性关闭的潜在介导物。其中一个序列AS3的表达模式与雄激素诱导的增殖关闭在时间上和DoseWise的表达都有很强的相关性。AS3基因位于染色体13 q12.3上，与BRCA2基因非常接近。AS3等位基因所在的染色体区域的缺失与包括前列腺癌在内的多种人类癌症有关，我们在两个稳定的细胞系中测试了AS3的生物学效应，其中一个表达正义AS3结构，另一个表达反义AS3结构，均受四环素调控。用含有四环素调控正义AS3结构的病毒粒子逆转录病毒感染S9细胞。在这些细胞中，正义AS3受到四环素的负调控。四环素停药后AS3mRNA和蛋白表达增加。四环素调控的AS3的表达抑制了细胞的增殖。用含有四环素调控的反义AS3结构的病毒粒子逆转录病毒感染A4细胞，载体驱动的反义AS3表达阻断雄激素诱导的内源性AS3mRNA的诱导，并阻断雄激素对细胞增殖的抑制作用。四环素调控的空载体对照表达对细胞增殖无影响。这些实验有力地表明，AS3是雄激素诱导的增殖阻断的中介物。
In the prostate gland of adult mammals, most epithelial cells are in a state of proliferative quiescence. Androgens regulate this effect by inducing cell cycle arrest in the G(0)/G(1) phase. Potential mediators of this androgen-induced proliferative shutoff were identified by means of subtracted cDNA libraries. The expression pattern of one of these sequences, AS3, strongly correlated with the expression of the androgen-induced proliferative shutoff both temporally and dosewise. The AS3 gene is located on chromosome 13 q12.3, in close proximity to the BRCA2 gene. The loss of chromosomal regions where AS3 alleles are located correlates with various human cancers, including prostate, The biological effect of AS3 was tested in two stable cell lines, one expressing sense and another expressing antisense AS3 constructs, both under tetracycline regulation. S9 cells were obtained by retroviral infection with virions containing a tetracycline-regulated sense AS3 construct. In these cells, sense AS3 was negatively regulated by tetracycline. Tetracycline withdrawal increased the expression of AS3 mRNA and protein. The expression of tetracycline-regulated AS3 resulted in inhibition of cell proliferation. A4 cells were obtained by retroviral infection with virions containing a tetracycline-regulated antisense AS3 construct, Vector-driven expression of antisense-AS3 blocked the induction of androgen-induced endogenous AS3 mRNA and blocked the inhibitory effect of androgens on cell proliferation. Tetracycline-regulated expression of the empty vector control had no effect on cell proliferation. These experiments strongly suggest that AS3 is a mediator of the androgen-induced proliferative shutoff.