He Xiaoqing, Despeaux Emily, Stueckle Todd A, Chi Alexander, Castranova Vincent, Dinu Cerasela Zoica, Wang Liying, Rojanasakul Yon
Department of Pharmaceutical Sciences, West Virginia University, Morgantown, West Virginia;
Department of Pharmaceutical Sciences, West Virginia University, Morgantown, West Virginia; HELD, National Institute for Occupational Safety and Health, Morgantown, West Virginia;
Am J Physiol Lung Cell Mol Physiol. 2016 Sep 1;311(3):L538-49. doi: 10.1152/ajplung.00139.2016. Epub 2016 Jul 15.
Carbon nanotubes (CNTs) have been likened to asbestos in terms of morphology and toxicity. CNT exposure can lead to pulmonary fibrosis and promotion of tumorigenesis. However, the mechanisms underlying CNT-induced carcinogenesis are not well defined. Mesothelin (MSLN) is overexpressed in many human tumors, including mesotheliomas and pancreatic and ovarian carcinomas. In this study, the role of MSLN in the carcinogenic transformation of human bronchial epithelial cells chronically exposed to single-walled CNT (BSW) was investigated. MSLN overexpression was found in human lung tumors, lung cancer cell lines, and BSW cells. The functional role of MSLN in the BSW cells was then investigated by using stably transfected MSLN knockdown (BSW shMSLN) cells. MSLN knockdown resulted in significantly decreased invasion, migration, colonies on soft agar, and tumor sphere formation. In vivo, BSW shMSLN cells formed smaller primary tumors and less metastases. The mechanism by which MSLN contributes to these more aggressive behaviors was investigated by using ingenuity pathway analysis, which predicted that increased MSLN could induce cyclin E expression. We found that BSW shMSLN cells had decreased cyclin E, and their proliferation rate was reverted to nearly that of untransformed cells. Cell cycle analysis showed that the BSW shMSLN cells had an increased G2 population and a decreased S phase population, which is consistent with the decreased rate of proliferation. Together, our results indicate a novel role of MSLN in the malignant transformation of bronchial epithelial cells following CNT exposure, suggesting its utility as a potential biomarker and drug target for CNT-induced malignancies.
碳纳米管(CNT)在形态和毒性方面被比作石棉。接触CNT可导致肺纤维化并促进肿瘤发生。然而,CNT诱导致癌作用的潜在机制尚未明确。间皮素(MSLN)在许多人类肿瘤中过度表达,包括间皮瘤、胰腺癌和卵巢癌。在本研究中,研究了MSLN在长期暴露于单壁CNT(BSW)的人支气管上皮细胞致癌转化中的作用。在人肺肿瘤、肺癌细胞系和BSW细胞中发现了MSLN的过度表达。然后通过使用稳定转染的MSLN敲低(BSW shMSLN)细胞研究了MSLN在BSW细胞中的功能作用。MSLN敲低导致侵袭、迁移、软琼脂上的集落形成和肿瘤球形成显著减少。在体内,BSW shMSLN细胞形成的原发性肿瘤较小且转移较少。通过 Ingenuity 通路分析研究了MSLN促成这些更具侵袭性的行为的机制,该分析预测MSLN增加可诱导细胞周期蛋白E表达。我们发现BSW shMSLN细胞的细胞周期蛋白E减少,其增殖率恢复到接近未转化细胞的水平。细胞周期分析表明,BSW shMSLN细胞的G2期细胞增多,S期细胞减少,这与增殖率降低一致。总之,我们的结果表明MSLN在CNT暴露后支气管上皮细胞的恶性转化中具有新作用,提示其作为CNT诱导恶性肿瘤的潜在生物标志物和药物靶点的效用。