Department of Respiratory and Critical Care Medicine, First Affiliated Hospital of Gannan Medical University, Ganzhou City, Jiangxi Province, China.
Department of Oncology, First Affiliated Hospital of Gannan Medical University, Ganzhou City, Jiangxi Province, China.
J Biochem Mol Toxicol. 2024 Aug;38(8):e23801. doi: 10.1002/jbt.23801.
Lung cancer (LC) is a major inducer of cancer-related death. We aim to reveal the effect of Calsequestrin2 (CASQ2) on macrophage polarization and Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in LC. Hub genes were determined from protein-protein interaction networks based on GSE21933 and GSE1987 data sets using bioinformatic analysis. Expression of hub genes was verified by real-time quantitative polymerase chain reaction (RT-qPCR). Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, wound-healing, colony formation, and transwell assays were performed to assess the impact of CASQ2 on LC cells. A xenograft mouse model was evaluated using hematoxylin-eosin, immunohistochemistry, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining to investigate the effect of CASQ2 on LC. The role of CASQ2 in regulating macrophage polarization and JAK/STAT pathway was evaluated by western blot andRT-qPCR. We screened out 155 common differentially expressed genes in GSE21933 and GSE1987 data sets. Myomesin-2, tyrosine kinase, sex determining region Y-box 2, platelet and endothelial cell adhesion molecule 1, matrix metallopeptidase 9, claudin-5, caveolin-1, CASQ2, recombinant ATPase, Ca++ transporting, cardiac muscle, slow twitch 2 (ATP2A2), and ankyrin repeat domain 1 were identified as the hub genes with high prediction value. CASQ2 was selected as a pivotal regulator of LC. In vitro experiments and xenograft models revealed that CASQ2 overexpression suppressed proliferation, colony formation, migration, invasion of LC cells, and tumor growth in vivo. Additionally, overexpression of CASQ2 promoted the expression of M1 macrophage markers (cluster of differentiation 80 [CD80], interleukin [IL]-12, inducible nitric oxide synthase [iNOS]), while decreasing the expression of M2 macrophage markers (CD163, IL-10, Arg1) in tumor-associated macrophages and xenograft tissues. Finally, we found that overexpression of CASQ2 inhibited JAK/STAT pathway. CASQ2 is a novel biomarker, which can alleviate LC via inhibiting M2 tumor-associated macrophage polarization and JAK/STAT pathway.
肺癌（LC）是癌症相关死亡的主要诱因。我们旨在揭示钙结合蛋白 2（CASQ2）对 LC 中巨噬细胞极化和 Janus 激酶/信号转导和转录激活因子（JAK/STAT）通路的影响。基于蛋白质-蛋白质相互作用网络，使用生物信息学分析从 GSE21933 和 GSE1987 数据集确定了枢纽基因。使用实时定量聚合酶链反应（RT-qPCR）验证了枢纽基因的表达。通过细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷、划痕愈合、集落形成和 Transwell 测定评估 CASQ2 对 LC 细胞的影响。使用苏木精-伊红、免疫组织化学和末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记染色评估 CASQ2 对 LC 的影响。通过 Western blot 和 RT-qPCR 评估 CASQ2 对巨噬细胞极化和 JAK/STAT 通路的调节作用。在 GSE21933 和 GSE1987 数据集中共筛选出 155 个共同差异表达基因。肌球蛋白-2、酪氨酸激酶、性别决定区 Y 盒 2、血小板和内皮细胞黏附分子 1、基质金属蛋白酶 9、闭合蛋白-5、窖蛋白-1、CASQ2、重组 ATP 酶，Ca++转运，心肌，慢肌 2（ATP2A2）和锚蛋白重复域 1 被鉴定为具有高预测价值的枢纽基因。CASQ2 被选为 LC 的关键调节剂。体外实验和异种移植模型表明，CASQ2 过表达抑制 LC 细胞的增殖、集落形成、迁移和侵袭，以及体内肿瘤生长。此外，CASQ2 过表达促进肿瘤相关巨噬细胞和异种移植组织中 M1 巨噬细胞标志物（分化群 80 [CD80]、白细胞介素 [IL]-12、诱导型一氧化氮合酶 [iNOS]）的表达，同时降低 M2 巨噬细胞标志物（CD163、IL-10、Arg1）的表达。最后，我们发现 CASQ2 过表达抑制 JAK/STAT 通路。CASQ2 是一种新型生物标志物，可通过抑制 M2 肿瘤相关巨噬细胞极化和 JAK/STAT 通路缓解 LC。
