Woolery Kamisha T, Mohamed Mai, Linger Rebecca J, Dobrinski Kimberly P, Roman Jesse, Kruk Patricia A
Department of Pathology & Cell Biology, University of South Florida, Tampa, FL 33612, USA.
Department of Medicine, University of Louisville, Louisville, KY 40202, USA.
Biomed Res Int. 2015;2015:652017. doi: 10.1155/2015/652017. Epub 2015 Aug 19.
Familial history remains the strongest risk factor for developing ovarian cancer (OC) and is associated with germline BRCA1 mutations, such as the 185delAG founder mutation. We sought to determine whether normal human ovarian surface epithelial (OSE) cells expressing the BRCA1 185delAG mutant, BRAT, could promote an inflammatory phenotype by investigating its impact on expression of the proinflammatory cytokine, Interleukin-1β (IL-1β). Cultured OSE cells with and without BRAT were analyzed for differential target gene expression by real-time PCR, western blot, ELISA, luciferase reporter, and siRNA assays. We found that BRAT cells expressed increased cellular and secreted levels of active IL-1β. BRAT-expressing OSE cells exhibited 3-fold enhanced IL-1β mRNA expression, transcriptionally regulated, in part, through CREB sites within the (-1800) to (-900) region of its promoter. In addition to transcriptional regulation, BRAT-mediated IL-1β expression appears dualistic through enhanced inflammasome-mediated caspase-1 cleavage and activation of IL-1β. Further investigation is warranted to elucidate the molecular mechanism(s) of BRAT-mediated IL-1β expression since increased IL-1β expression may represent an early step contributing to OC.
家族病史仍然是患卵巢癌(OC)的最强风险因素,并且与种系BRCA1突变相关,比如185delAG始祖突变。我们试图通过研究表达BRCA1 185delAG突变体BRAT的正常人类卵巢表面上皮(OSE)细胞对促炎细胞因子白细胞介素-1β(IL-1β)表达的影响,来确定其是否能促进炎症表型。通过实时PCR、蛋白质免疫印迹、酶联免疫吸附测定、荧光素酶报告基因和小干扰RNA测定,对培养的有和没有BRAT的OSE细胞进行差异靶基因表达分析。我们发现BRAT细胞中活性IL-1β的细胞内水平和分泌水平均升高。表达BRAT的OSE细胞表现出IL-1β mRNA表达增强3倍,这部分是通过其启动子(-1800)至(-900)区域内的CREB位点进行转录调控的。除了转录调控外,BRAT介导的IL-1β表达似乎具有双重性,即通过增强炎性小体介导的caspase-1切割和IL-1β的激活来实现。由于IL-1β表达增加可能代表了导致OC的早期步骤,因此有必要进一步研究以阐明BRAT介导的IL-1β表达的分子机制。