Identification of ATF-3, caveolin-1, DLC-1, and NM23-H2 as putative antitumorigenic, progesterone-regulated genes for ovarian cancer cells by gene profiling
Identification of ATF-3, caveolin-1, DLC-1, and NM23-H2 as putative antitumorigenic, progesterone-regulated genes for ovarian cancer cells by gene profiling
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DOI：
10.1038/sj.onc.1207991
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发表时间：
2005-03-03
期刊：
影响因子：
8
通讯作者：
Ho, SM
中科院分区：
文献类型：
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作者：
Syed, V;Mukherjee, K;Ho, SM
虽然孕酮（P4）被认为可以预防卵巢癌（OCa），但对其作用机制知之甚少。本研究的目的是鉴定具有抗OCa作用的P4调节基因。三个永生化的非致瘤性人卵巢表面上皮（HOSE）细胞系和三个OCa（OVCA）细胞系进行5天的P4处理。转录亲。用含有大约2400个已知基因的cDNA微阵列进行测序，以鉴定（1）与HOSE细胞系相比，OVCA细胞系中表达持续下调的基因，和（2）经P4处理后，OCa细胞系中表达恢复的基因。从所选的候选物中，选择转录激活因子-3（ATF-3）、小窝蛋白-1、肝癌中缺失的小窝蛋白-1（DLC-1）和非转移性克隆23（NM 23-H2）用于基于它们先前报道的作为肿瘤抑制剂或凋亡诱导剂的作用的事后功能研究。半定量RT-PCR分析证实，与HOSE细胞相比，OVCA细胞中这些基因的转录丢失或减少，并且在P4处理后它们上调。激素特异性通过用孕酮拮抗剂RU 38486的阻断实验证明。小窝蛋白-1，DLC-1和NM 23-H2的异位表达引起OVCA细胞培养物的生长抑制，但在HOSE细胞培养物中不引起生长抑制，而ATF-3的强制表达抑制两者的生长。AFT-3的过表达也增强了HOSE和OVCA细胞中caspase-3的活性，而小窝蛋白-1和DLC-1的异位表达仅激活了OCa细胞中的这种酶。相反，NM 23-H2过表达在激活caspase-3方面无效。OCa细胞中四种基因中任何一种的过表达都降低了软琼脂集落形成和细胞侵袭力。总之，我们已经确定了四个新的P4调节，抗肿瘤基因的OCa。然而，它们的作用模式显著不同; ATF-3主要用作凋亡诱导剂，NM 23-H2作为细胞运动的抑制剂，而小窝蛋白-1和DLC-1表现出经典肿瘤抑制剂的特征。据我们所知，除了NM 23-H2，这是第一个报告连接P4的这些肿瘤抑制/促凋亡基因的调节，这可能是未来的治疗目标。
Although progesterone (P4) has been implicated to offer protection against ovarian cancer (OCa), little is known of its mechanism of action. The goal of this study was to identify P4-regulated genes that have anti-OCa action. Three immortalized nontumorigenic human ovarian surface epithelial ( HOSE) cell lines and three OCa (OVCA) cell lines were subjected to 5 days of P4 treatment. Transcriptional pro. ling with a cDNA microarray containing approximate to2400 known genes was used to identify genes (1) whose expression was consistently downregulated in OVCA cell lines compared to HOSE cell lines, and ( 2) whose expression was restored in OCa cell lines by P4 treatment. From the candidates selected, activating transcription factor-3 (ATF-3), caveolin-1, deleted in liver cancer-1 (DLC-1), and nonmetastatic clone 23 ( NM23-H2) were chosen for post hoc functional studies based on their previously reported action as tumor suppressors or apoptosis inducers. Semiquantitative RT-PCR analyses confirmed loss of or reduced transcription of these genes in OVCA cells when compared to HOSE cells and their upregulation following P4 treatment. Hormonal specificity was demonstrated by blockade experiments with a progestin antagonist RU 38486. Ectopic expression of caveolin-1, DLC-1, and NM23-H2 caused growth inhibition in OVCA cell cultures, but not in HOSE cell cultures, while forced expression of ATF-3 suppressed growth in both. Overexpression of AFT-3 also enhanced caspase-3 activity in both HOSE and OVCA cells, whereas ectopic expression of caveolin-1 and DLC-1 only activated this enzyme in OCa cells. In contrast, NM23-H2 overexpression was ineffective in activating caspase-3. Overexpression of any of the four genes in OCa cells reduced soft-agar colony formation and cell invasiveness. Taken together, we have identified four new P4-regulated, antitumor genes forOCa. However, their modes of action differ significantly; ATF-3 primarily functions as an apoptosis inducer, NM23-H2 as a suppressor of cell motility, and caveolin-1 and DLC-1 exhibiting features of classical tumor suppressors. To the best of our knowledge, except for NM23-H2, this is the first report linking P4 to the regulation of these tumor suppressor/proapoptotic genes, which could serve as future therapeutic targets.