接触石棉样氟直闪石纤维后生物功能指标的体外研究

Pugnaloni A, Lucarini G, GiantomassI F, Lombardo L, Capella S, Belluso E, Zizzi A, Panico A M, Biagini G, Cardile V

Dipartimento di Patologia Molecolare e Terapie Innovative-Istologia, Università Politecnica delle Marche, Via Tronto, 10/A, 60020, Torrette, Ancona, Italy,

Cell Mol Biol (Noisy-le-grand). 2007;53 Suppl:OL965-80.

The in vitro biological response to fluoro-edenite (FE) fibres, an asbestos-like amphibole, was evaluated in lung alveolar epithelial A549, mesothelial MeT-5A and monocyte-macrophage J774 cell lines. The mineral has been found in the vicinity of the town of Biancavilla (Catania, Sicily), where an abnormal incidence of mesothelioma has been documented. Cell motility, distribution of polymerized actin, and synthesis of vascular endothelial growth factor (VEGF) and of beta-catenin, critical parameters for tumour development, progression and survival, were investigated in A549 and MeT-5A cells exposed to 50 microg/ml FE fibres for 24 hr and 48 hr. The levels of cyclooxygenase (COX-2) and prostaglandin (PGE2), two molecules involved in cancer pathogenesis by affecting mitogenesis, cell adhesion, immune surveillance and apoptosis, were measured in J774 cells treated with FE fibres under the same experimental conditions. Finally, FE fibres were studied by SEM and EDS analysis to investigate their chemical composition. Exposure of A549 and MeT-5A cells to FE fibres affected differentially phalloidin-stained cytoplasmic F-actin networks, cell motility and VEGF and beta-catenin expression according to the different sensitivity of the two cell lines. In J774 cells it induced a significant increase in COX-2 expression, as assessed by Western blot analysis, and in the concentration of PGE2, measured in culture media by ELISA. SEM-EDS investigations demonstrated two types of FE fibres, edenite and fluoro-edenite, differing in chemical composition and both recognizable as calcic amphiboles. Fibre width ranged from less than 1 microm (prevalently 0.5 microm) to 2-3 microm (edenite) up to several microm (fluoro-edenite); length ranged from about 6 to 80 microm (edenite) up to some hundred microm (fluoro-edenite). Results provide convincing evidence that FE fibres are capable of inducing in vitro functional modifications in a number of parameters with crucial roles in cancer development and progression. Inhaled FE fibres have the potential to induce mesothelioma, even though their ability to penetrate lung alveoli depends on their aerodynamic diameter.

在肺泡上皮A549、间皮MeT-5A和单核巨噬细胞J774细胞系中评估了对氟直闪石（FE）纤维（一种类似石棉的闪石）的体外生物学反应。在比安卡维拉镇（西西里岛卡塔尼亚）附近发现了这种矿物质，在那里间皮瘤的发病率异常高。在暴露于50微克/毫升FE纤维24小时和48小时的A549和MeT-5A细胞中，研究了细胞运动性、聚合肌动蛋白的分布以及血管内皮生长因子（VEGF）和β-连环蛋白的合成，这些是肿瘤发生、发展和存活的关键参数。在相同实验条件下，用FE纤维处理的J774细胞中，测量了环氧化酶（COX-2）和前列腺素（PGE2）的水平，这两种分子通过影响有丝分裂、细胞粘附、免疫监视和细胞凋亡参与癌症发病机制。最后，通过扫描电子显微镜（SEM）和能谱分析（EDS）研究FE纤维以调查其化学成分。根据两种细胞系的不同敏感性，A549和MeT-5A细胞暴露于FE纤维会差异性地影响鬼笔环肽染色的细胞质F-肌动蛋白网络、细胞运动性以及VEGF和β-连环蛋白的表达。在J774细胞中，通过蛋白质印迹分析评估发现它会使COX-2表达显著增加，并且通过酶联免疫吸附测定（ELISA）在培养基中测量发现PGE2浓度升高。扫描电子显微镜-能谱分析（SEM-EDS）研究表明存在两种类型的FE纤维，即直闪石和氟直闪石，它们的化学成分不同，且都可识别为钙闪石。纤维宽度范围从小于1微米（主要为0.5微米）到2 - 3微米（直闪石）直至几微米（氟直闪石）；长度范围从约6到80微米（直闪石）直至几百微米（氟直闪石）。结果提供了令人信服的证据，表明FE纤维能够在体外诱导一些对癌症发展和进展起关键作用的参数发生功能改变。吸入的FE纤维有可能诱发间皮瘤，尽管它们穿透肺泡的能力取决于其空气动力学直径。