Thériault Brigitte L, Basavarajappa Halesha D, Lim Harvey, Pajovic Sanja, Gallie Brenda L, Corson Timothy W
Campbell Family Cancer Research Institute, Princess Margaret Cancer Centre, Toronto, Ontario, Canada.
Eugene and Marilyn Glick Eye Institute, Department of Ophthalmology, and Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
PLoS One. 2014 Mar 13;9(3):e91540. doi: 10.1371/journal.pone.0091540. eCollection 2014.
KIF14 (kinesin family member 14) is a mitotic kinesin and an important oncogene in several cancers. Tumor KIF14 expression levels are independently predictive of poor outcome, and in cancer cells KIF14 can modulate metastatic behavior by maintaining appropriate levels of cell adhesion and migration proteins at the cell membrane. Thus KIF14 is an exciting potential therapeutic target. Understanding KIF14's regulation in cancer cells is crucial to the development of effective and selective therapies to block its tumorigenic function(s). We previously determined that close to 30% of serous ovarian cancers (OvCa tumors) exhibit low-level genomic gain, indicating one mechanism of KIF14 overexpression in tumors. We now report on transcriptional and epigenetic regulation of KIF14. Through promoter deletion analyses, we identified one cis-regulatory region containing binding sites for Sp1, HSF1 and YY1. siRNA-mediated knockdown of these transcription factors demonstrated endogenous regulation of KIF14 overexpression by Sp1 and YY1, but not HSF1. ChIP experiments confirmed an enrichment of both Sp1 and YY1 binding to the endogenous KIF14 promoter in OvCa cell lines with high KIF14 expression. A strong correlation was seen in primary serous OvCa tumors between Sp1, YY1 and KIF14 expression, further evidence that these transcription factors are important players in KIF14 overexpression. Hypomethylation patterns were observed in primary serous OvCa tumors, suggesting a minor role for promoter methylation in the control of KIF14 gene expression. miRNA expression analysis determined that miR-93, miR-144 and miR-382 had significantly lower levels of expression in primary serous OvCa tumors than normal tissues; treatment of an OvCa cell line with miRNA mimics and inhibitors specifically modulated KIF14 mRNA levels, pointing to potential novel mechanisms of KIF14 overexpression in primary tumors. Our findings reveal multiple mechanisms of KIF14 upregulation in cancer cells, offering new targets for therapeutic interventions to reduce KIF14 in tumors, aiming at improved prognosis.
KIF14(驱动蛋白家族成员14)是一种有丝分裂驱动蛋白,也是多种癌症中的重要致癌基因。肿瘤中KIF14的表达水平可独立预测不良预后,在癌细胞中,KIF14可通过维持细胞膜上细胞黏附蛋白和迁移蛋白的适当水平来调节转移行为。因此,KIF14是一个令人兴奋的潜在治疗靶点。了解KIF14在癌细胞中的调控对于开发有效且有选择性的疗法以阻断其致瘤功能至关重要。我们之前确定,近30%的浆液性卵巢癌(OvCa肿瘤)表现出低水平的基因组扩增,这表明肿瘤中KIF14过表达的一种机制。我们现在报告KIF14的转录和表观遗传调控。通过启动子缺失分析,我们鉴定出一个顺式调控区域,其含有Sp1、HSF1和YY1的结合位点。siRNA介导的这些转录因子敲低表明,Sp1和YY1对内源性KIF14过表达有调控作用,而HSF1没有。染色质免疫沉淀实验证实,在KIF14高表达的OvCa细胞系中,Sp1和YY1均富集于内源性KIF14启动子。在原发性浆液性OvCa肿瘤中,Sp1、YY1和KIF14表达之间存在很强的相关性,进一步证明这些转录因子是KIF14过表达的重要因素。在原发性浆液性OvCa肿瘤中观察到低甲基化模式,表明启动子甲基化在KIF14基因表达控制中的作用较小。miRNA表达分析确定,miR-93、miR-·144和miR-382在原发性浆液性OvCa肿瘤中的表达水平明显低于正常组织;用miRNA模拟物和抑制剂处理OvCa细胞系可特异性调节KIF14 mRNA水平,这表明原发性肿瘤中KIF14过表达可能存在新机制。我们的研究结果揭示了癌细胞中KIF14上调的多种机制,为旨在改善预后的降低肿瘤中KIF14水平的治疗干预提供了新靶点。