Yamamoto Kazuo, Seike Masahiro, Takeuchi Susumu, Soeno Chie, Miyanaga Akihiko, Noro Rintaro, Minegishi Yuji, Kubota Kaoru, Gemma Akihiko
Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo 113-8603, Japan.
Oncol Rep. 2014 Dec;32(6):2365-72. doi: 10.3892/or.2014.3481. Epub 2014 Sep 16.
Malignant pleural mesothelioma (MPM) is a rapidly fatal malignancy that is increasing in incidence in Japan. In this study, we performed gene and microRNA (miRNA) expression profiling to identify novel therapeutic targets in MPM cells. Based on relative sensitivities to pemetrexed (PEM) and the histone deacetylase (HDAC) inhibitor, vorinostat (SAHA), 211H cells were determined to be the only sensitive MPM cell line out of the 6 tested. On the same series of cell lines, we performed whole genome transcriptomic profiling via DNA microarrays and pathway analysis of the derived data. Of particular note, IL-18 gene expression levels were significantly higher in the cell lines that were either drug resistant or displayed intermediate sensitivity, compared to the sensitive 211H cell line. Pathway analysis revealed IL-18 as an important gene associated with drug sensitivity of MPM cells. A relationship between IL-18 overexpression and drug resistance was also observed following targeted assessment of 10 cytokine genes using quantitative RT-PCR. miRNA expression profiles were evaluated in the MPM cell line panel in order to discern the mechanism of IL-18 induction in the drug-resistant lines. We found that miR-379 and miR-411 belonged to the same cluster of miRNAs located on chromosome 14q32 that commonly target the IL-18 gene. Luciferase reporter assays revealed that miR-379 and miR-411 directly target the IL-18 gene. Introduction of miR-379 plus miR-411, as well as IL-18 silencing, significantly suppressed the invasive capacity of MESO1 cells in vitro. Furthermore, the use of either PEM or SAHA together with miR-379 plus miR-411 mimics mediated increased sensitivity to these drugs in MESO1 cells. These results suggest that the miR-379/411 cluster may provide new therapeutic opportunities for advanced MPM patients, depending on the nature of IL-18 gene expression.
恶性胸膜间皮瘤（MPM）是一种快速致死性恶性肿瘤，在日本其发病率正在上升。在本研究中，我们进行了基因和微小RNA（miRNA）表达谱分析，以确定MPM细胞中的新型治疗靶点。基于对培美曲塞（PEM）和组蛋白去乙酰化酶（HDAC）抑制剂伏立诺他（SAHA）的相对敏感性，在6个测试的MPM细胞系中，211H细胞系被确定为唯一敏感的细胞系。在同一系列细胞系上，我们通过DNA微阵列进行了全基因组转录组分析，并对所得数据进行了通路分析。特别值得注意的是，与敏感的211H细胞系相比，耐药或表现出中等敏感性的细胞系中白细胞介素-18（IL-18）基因表达水平显著更高。通路分析显示IL-18是与MPM细胞药物敏感性相关的重要基因。使用定量逆转录聚合酶链反应（RT-PCR）对10个细胞因子基因进行靶向评估后，也观察到IL-18过表达与耐药之间的关系。为了弄清耐药细胞系中IL-18诱导的机制，我们在MPM细胞系组中评估了miRNA表达谱。我们发现miR-379和miR-411属于位于14号染色体q32上的同一miRNA簇，它们共同靶向IL-18基因。荧光素酶报告基因检测显示miR-379和miR-411直接靶向IL-18基因。导入miR-379加miR-411以及沉默IL-18，在体外显著抑制了MESO1细胞的侵袭能力。此外，将PEM或SAHA与miR-379加miR-411模拟物联合使用，可介导MESO1细胞对这些药物的敏感性增加。这些结果表明，根据IL-?18基因表达的性质，miR-379/411簇可能为晚期MPM患者提供新的治疗机会。
