Division of Thoracic Surgery, Department of Medicine, Stanford University School of Medicine, Stanford, CA.
Department of Cardiothoracic Surgery, Department of Anesthesia, Department of Medicine, Stanford University School of Medicine, Stanford, CA.
Chest. 2013 Nov;144(5):1632-1643. doi: 10.1378/chest.12-2770.
We investigated microRNA expression profiles of malignant pleural mesothelioma (MPM) specimens to identify novel microRNA that are potentially involved in the oncogenic transformation of human pleural cells.
microRNA microarray transcriptional profiling studies of 25 MPM primary tumors were performed. We used normal pleural tissue from an unmatched patient cohort as normal comparators. To confirm microarray data, we used real-time quantitative polymerase chain reaction. Representative cell lines H513 and H2052 were used in functional analyses of miR-1.
In addition to several novel MPM-associated microRNAs, we observed that the expression level of miR-1 was significantly lower in tumors as compared with normal pleural specimens. Subsequently, pre-miR of miR-1 was introduced into MPM cell lines to overexpress this microRNA. Phenotypic changes of these altered cells were assayed. The cellular proliferation rate was significantly inhibited after overexpression of miR-1. Early and late apoptosis was increased markedly in miR-1-transfected cell lines. Taken together, these data suggested that overexpression of miR-1 induced apoptosis in these MPM cell lines, acting as a tumor suppressor. We confirmed our observations by assessing in the transduced MPM cells cell cycle-related, proapoptotic, and antiapoptotic genes, which all showed coordinated, significant changes characteristic of the apoptotic phenotype.
Further investigation and validation of our microRNA database of MPM may elucidate previously unrecognized molecular pathways and/or mechanisms by identifying novel microRNAs that are involved in malignant transformation. Our study has now found miR-1 to be one of these MPM-associated microRNAs, with potential pathogenic and therapeutic significance.
我们研究了恶性胸膜间皮瘤(MPM)标本的 microRNA 表达谱,以鉴定可能参与人胸膜细胞致癌转化的新型 microRNA。
对 25 例 MPM 原发性肿瘤进行了 microRNA 微阵列转录谱研究。我们使用来自不匹配患者队列的正常胸膜组织作为正常对照。为了验证微阵列数据,我们使用了实时定量聚合酶链反应。代表性细胞系 H513 和 H2052 用于 miR-1 的功能分析。
除了几种新型 MPM 相关的 microRNA 外,我们还观察到 miR-1 的表达水平在肿瘤组织中明显低于正常胸膜标本。随后,将 miR-1 的前体 miRNA 引入 MPM 细胞系中以过表达这种 microRNA。这些改变细胞的表型变化被检测。过表达 miR-1 后,细胞增殖率显著受到抑制。miR-1 转染细胞系中的早期和晚期凋亡明显增加。总的来说,这些数据表明,过表达 miR-1 诱导这些 MPM 细胞系凋亡,发挥肿瘤抑制作用。我们通过评估转导的 MPM 细胞中的细胞周期相关、促凋亡和抗凋亡基因,证实了我们的观察结果,这些基因都显示出与凋亡表型一致的协调、显著变化。
进一步研究和验证我们的 MPM microRNA 数据库可能阐明以前未被识别的分子途径和/或机制,通过鉴定可能参与恶性转化的新型 microRNA。我们的研究现在发现 miR-1 是这些 MPM 相关 microRNA 之一,具有潜在的致病和治疗意义。