Automated High-Content Screening for Compounds That Disassemble the Perinucleolar Compartment
Automated High-Content Screening for Compounds That Disassemble the Perinucleolar Compartment
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DOI：
10.1177/1087057109343120
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发表时间：
2009-10-01
影响因子：
--
通讯作者：
Huang, Sui
中科院分区：
文献类型：
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作者：
Norton, John T.;Titus, Steven A.;Huang, Sui
所有实体恶性肿瘤都有共同的特征，包括无限制的细胞增殖、逃避免疫调节和转移倾向。作者之前曾描述过，亚核结构（核周区室（PNC））与实体瘤癌细胞的转移表型相关。含有PNC的癌细胞的百分比（PNC患病率）指示体外和体内肿瘤的恶性程度，因此PNC患病率是反映肿瘤细胞群体中转移能力的标志物。虽然PNC的功能仍有待确定，但PNC高度富集小RNA和RNA结合蛋白。最初的化学生物学研究使用一组分解PNC的抗癌药物，揭示了结构与DNA的直接关联。因此，作为表型标志物的PNC流行率降低可用于鉴定靶向PNC维持所需的细胞过程的化合物，并因此用于阐明PNC功能的性质。在这里，作者报告了一种自动化高含量筛选测定的发展，该测定能够检测稳定表达定位于PNC的绿色荧光蛋白（GFP）融合蛋白的前列腺癌细胞（PC-3 M）中PNC的患病率。使用已知的PNC还原药物和非PNC还原细胞毒性药物优化测定。优化后，该测定的保真度用8284种化合物的集合进行了探测，并显示出稳健性，能够检测已知和新型的PNC还原化合物，使其成为首次报道的核结构微小变化的高含量表型筛选。（Journal of Biomolecular Screening 2009：1045-1053）
All solid malignancies share characteristic traits, including unlimited cellular proliferation, evasion of immune regulation, and the propensity to metastasize. The authors have previously described that a subnuclear structure, the perinucleolar compartment (PNC), is associated with the metastatic phenotype in solid tumor cancer cells. The percentage of cancer cells that contain PNCs (PNC prevalence) is indicative of the malignancy of a tumor both in vitro and in vivo, and thus PNC prevalence is a marker that reflects metastatic capability in a population of tumor cells. Although the function of the PNC remains to be determined, the PNC is highly enriched with small RNAs and RNA binding proteins. The initial chemical biology studies using a set of anticancer drugs that disassemble PNCs revealed a direct association of the structure with DNA. Therefore, PNC prevalence reduction as a phenotypic marker can be used to identify compounds that target cellular processes required for PNC maintenance and hence used to elucidate the nature of the PNC function. Here the authors report the development of an automated high-content screening assay that is capable of detecting PNC prevalence in prostate cancer cells (PC-3M) stably expressing a green fluorescent protein (GFP)-fusion protein that localizes to the PNC. The assay was optimized using known PNC-reducing drugs and non-PNC-reducing cytotoxic drugs. After optimization, the fidelity of the assay was probed with a collection of 8284 compounds and was shown to be robust and capable of detecting known and novel PNC-reducing compounds, making it the first reported high-content phenotypic screen for small changes in nuclear structure. (Journal of Biomolecular Screening 2009:1045-1053)