Yang Dan, Chen Chiyun, Xia Hailing, Chen Junqiang, Yu Min
School of Public Heath, Hangzhou Medical College, Hangzhou, Zhejiang, China.
Department of Pulmonary and Critical Care Medicine, Cixi People's Hospital, Cixi, Zhejiang, China.
J Appl Toxicol. 2023 Oct;43(10):1511-1521. doi: 10.1002/jat.4493. Epub 2023 May 10.
Asbestos is a fibrous silicate mineral exhibiting biopersistence and carcinogenic properties and contributes to mesothelioma. Despite the concept of gene-environmental interaction in pathogenesis of mesothelioma, the possible pathophysiological changes of mesothelial cells simultaneously with SET domain containing 2 (SETD2) loss and asbestos exposure remains obscure. Herein, CRISPR/Cas9-mediated SETD2 knockout Met-5A mesothelial cells (Met-5A ) were established and exposed with crocidolite, an amphibole asbestos. Cell viability of Met-5A appeared to dramatically decrease with ≥2.5 μg/cm crocidolite exposure as compared with Met-5A, although no cytotoxicity and apoptosis changes of Met-5A and Met-5A was evident with 1.25 μg/cm crocidolite exposure for 48 h. RNA sequencing uncovered top 50 differentially expressed genes (DEGs) between 1.25 μg/cm crocidolite exposed Met-5A (Cro-Met-5A ) and 1.25 μg/cm crocidolite exposed Met-5A (Cro-Met-5A), and ITGA4, THBS2, MYL7, RAC2, CADM1, and CLDN11 appeared to be the primary DEGs involved with adhesion in gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Cro-Met-5A had strong migration but mild adhesion behavior as compared with Cro-Met-5A. Additionally, crocidolite tended to increase migration of Met-5A but inhibited migration of Met-5A when compared with their corresponding cells without crocidolite exposure, although no further adhesion property changes was evident for both cells in response to crocidolite. Therefore, crocidolite may affect adhesion-related gene expression and modify adhesion and migration behavior for SETD2-depleted Met-5A, which could provide preliminary insight regarding the potential role of SETD2 in the cell behavior of asbestos-related malignant mesothelial cell.
石棉是一种纤维状硅酸盐矿物，具有生物持久性和致癌特性，可导致间皮瘤。尽管在间皮瘤发病机制中存在基因-环境相互作用的概念，但在SET结构域包含蛋白2（SETD2）缺失和接触石棉的同时，间皮细胞可能发生的病理生理变化仍不清楚。在此，我们建立了CRISPR/Cas9介导的SETD2基因敲除的Met-5A间皮细胞（Met-5A），并用阳起石石棉（一种闪石类石棉）进行处理。与Met-5A相比，当阳起石暴露量≥2.5μg/cm时，Met-5A的细胞活力似乎显著下降，不过在1.25μg/cm阳起石暴露48小时时，Met-5A和Met-5A均未出现明显的细胞毒性和凋亡变化。RNA测序揭示了在1.25μg/cm阳起石处理的Met-5A（Cro-Met-5A）和1.25μg/cm阳起石处理的Met-5A（Cro-Met-5A）之间排名前50的差异表达基因（DEG），在基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析中，整合素α4（ITGA4）、血小板反应蛋白2（THBS2）、肌球蛋白轻链7（MYL7）、RAC家族小GTP酶2（RAC2）、细胞粘附分子1（CADM1）和紧密连接蛋白11（CLDN11）似乎是与粘附相关的主要DEG。与Cro-Met-5A相比，Cro-Met-5A具有较强的迁移能力但粘附能力较弱。此外，与未接触阳起石的相应细胞相比，阳起石倾向于增加Met-5A的迁移能力，但抑制Met-5A的迁移能力，尽管两种细胞在接触阳起石后均未出现进一步明显的粘附特性变化。因此，阳起石可能会影响与粘附相关的基因表达，并改变SETD2缺失的Met-5A的粘附和迁移行为，这可为SETD2在石棉相关恶性间皮细胞的细胞行为中的潜在作用提供初步见解。
