Delineation of the molecular basis for selenium-induced growth arrest in human prostate cancer cells by oligonucleotide array.
Delineation of the molecular basis for selenium-induced growth arrest in human prostate cancer cells by oligonucleotide array.
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发表时间：
2003
期刊：
影响因子：
11.2
通讯作者：
Yan Dong;Haitao Zhang;L. Hawthorn;H. Ganther;C. Ip
中科院分区：
文献类型：
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作者：
Yan Dong;Haitao Zhang;L. Hawthorn;H. Ganther;C. Ip
尽管硒对人类前列腺癌的干预越来越感兴趣，但目前关于硒作用的分子机制的信息很少。我们过去的研究表明，甲基亚硒酸（MSA）是研究硒的体外抗癌作用的一种很好的试剂。本研究旨在研究MSA在PC-3人前列腺癌细胞中的细胞和分子效应。暴露于生理浓度的MSA后，这些细胞表现出剂量和时间依赖性的生长抑制。MSA在多个过渡点延缓细胞周期进程，而不改变细胞周期不同阶段的细胞比例。膜联蛋白V和碘化丙啶标记的细胞的流式细胞仪分析表明，MSA的细胞凋亡的显着诱导。然后应用Affytelium人类基因组U95 A芯片进行阵列分析，以分析可能介导硒效应的基因表达变化。使用同步化细胞在时间过程实验（在12、24、36和48 h）中进行基因分析。大量具有不同生物学功能的潜在硒响应基因被鉴定出来。这些基因分为12个不同的动力学模式的调制MSA的集群。选择已知与细胞周期调控密切相关的10个基因的表达变化，通过Western分析进行验证，以确定阵列数据的可靠性。基于这些确认实验，获得了70%的一致率。阵列数据使我们能够专注于潜在关键基因的作用（例如，GADD 153、CHK 2、p21（WAF 1）、细胞周期蛋白A、CDK 1和DHFR）可能是MSA阻碍细胞周期进展的靶点。这些数据还为硒的新生物学效应提供了有价值的见解，例如抑制细胞侵袭，DNA修复和刺激转化生长因子β信号传导。本研究表明，全基因组分析的效用，阐明硒化学预防的机制。
Despite the growing interest in selenium intervention of prostate cancer in humans, scanty information is currently available on the molecular mechanism of selenium action. Our past research indicated that methylseleninic acid (MSA) is an excellent reagent for investigating the anticancer effect of selenium in vitro. The present study was designed to examine the cellular and molecular effects of MSA in PC-3 human prostate cancer cells. After exposure to physiological concentrations of MSA, these cells exhibited a dose- and time-dependent inhibition of growth. MSA retarded cell cycle progression at multiple transition points without changing the proportion of cells in different phases of the cell cycle. Flow cytometric analysis of annexin V- and propidium iodide-labeled cells showed a marked induction of apoptosis by MSA. Array analysis with the Affymetrix human genome U95A chip was then applied to profile the gene expression changes that might mediate the effects of selenium. Gene profiling was done in a time course experiment (at 12, 24, 36, and 48 h) using synchronized cells. A large number of potential selenium-responsive genes with diverse biological functions were identified. These genes fell into 12 clusters of distinct kinetics pattern of modulation by MSA. The expression changes of 10 genes known to be critically involved in cell cycle regulation were selected for verification by Western analysis to determine the reliability of the array data. An agreement rate of 70% was obtained based on these confirmation experiments. The array data enabled us to focus on the role of potential key genes (e.g., GADD153, CHK2, p21(WAF1), cyclin A, CDK1, and DHFR) that might be targets of MSA in impeding cell cycle progression. The data also provide valuable insights into novel biological effects of selenium, such as inhibition of cell invasion, DNA repair, and stimulation of transforming growth factor beta signaling. The present study demonstrates the utility of a genome-wide analysis to elucidate the mechanism of selenium chemoprevention.