Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, United States of America.
PLoS One. 2012;7(10):e47086. doi: 10.1371/journal.pone.0047086. Epub 2012 Oct 9.
Targeted therapies have been used to combat many tumor types; however, few have effectively improved the overall survival in women with epithelial ovarian cancer, begging for a better understanding of this deadly disease and identification of essential drivers of tumorigenesis that can be targeted effectively. Therefore, we used a loss-of-function screening approach to help identify molecular vulnerabilities that may represent key points of therapeutic intervention. We employed an unbiased high-throughput lethality screen using a 24,088 siRNA library targeting over 6,000 druggable genes and studied their effects on growth and/or survival of epithelial ovarian cancer (EOC) cell lines. The top 300 "hits" affecting the viability of A1847 cells were rescreened across additional EOC cell lines and non-tumorigenic, human immortalized ovarian epithelial cell lines. Fifty-three gene candidates were found to exhibit effects in all tumorigenic cell lines tested. Extensive validation of these hits refined the list to four high quality candidates (HSPA5, NDC80, NUF2, and PTN). Mechanistic studies show that silencing of three genes leads to increased apoptosis, while HSPA5 silencing appears to alter cell growth through G1 cell cycle arrest. Furthermore, two independent gene expression studies show that NDC80, NUF2 and PTN were significantly aberrantly overexpressed in serous adenocarcinomas. Overall, our functional genomics results integrated with the genomics data provide an important unbiased avenue towards the identification of prospective therapeutic targets for drug discovery, which is an urgent and unmet clinical need for ovarian cancer.
靶向治疗已被用于治疗多种肿瘤类型；然而，很少有治疗方法能有效提高上皮性卵巢癌患者的总生存率，这就需要我们更好地了解这种致命疾病，并确定可有效靶向的肿瘤发生的关键驱动因素。因此，我们使用了一种功能丧失筛选方法，以帮助识别可能代表治疗干预关键点的分子脆弱性。我们使用了一种无偏见的高通量致死性筛选方法，使用了针对超过 6000 个可用药基因的 24088 个 siRNA 文库，并研究了它们对上皮性卵巢癌（EOC）细胞系生长和/或存活的影响。对 A1847 细胞活力有影响的前 300 个“命中”靶点在其他 EOC 细胞系和非致瘤性的人永生化卵巢上皮细胞系中进行了重新筛选。发现 53 个候选基因对所有测试的致瘤细胞系均有影响。对这些命中靶点进行了广泛验证，将候选基因列表缩小到四个高质量候选基因（HSPA5、NDC80、NUF2 和 PTN）。机制研究表明，沉默三个基因会导致细胞凋亡增加，而 HSPA5 沉默似乎通过 G1 细胞周期阻滞改变细胞生长。此外，两项独立的基因表达研究表明，NDC80、NUF2 和 PTN 在浆液性腺癌中显著过表达。总的来说，我们的功能基因组学结果与基因组学数据相结合，为药物发现提供了一个重要的、无偏见的途径，以确定潜在的治疗靶点，这是卵巢癌的一个迫切和未满足的临床需求。
