PO.IM02.01 · 免疫学
LPS通过钙库操纵性钙内流诱导超氧阴离子依赖性巨噬细胞胞外诱捕网
LPS-induced superoxide anion-dependent macrophage extracellular traps by store-operated calcium entry
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
巨噬细胞胞外诱捕网（METs）参与肿瘤微环境中的慢性炎症，通过坏死性凋亡诱导的细胞外基质降解促进癌症进展。本研究探讨钙库操纵性钙内流（SOCE）在钙诱导的MET形成中的作用，该过程可放大巨噬细胞中的促肿瘤炎症反应。用脂多糖（LPS）或佛波醇酯（PMA）刺激小鼠巨噬细胞RAW264.7和人单核细胞U937，同时预先或不预先使用Ca2+螯合剂BAPTA-AM、机械敏感性Ca2+通道抑制剂GdCl3、SOCE抑制剂SKF96365或YM58483，或超氧阴离子清除剂百里醌（TQ）进行处理。经洋地黄皂苷通透化后，通过Lamin B免疫荧光染色鉴定发生METosis的细胞。LPS和PMA触发SOCE介导的Ca2+内流，导致胞质内Ca2+水平升高，从而激活NADPH氧化酶（NOX）产生超氧阴离子，最终导致MET形成。药理学抑制SOCE减弱了Ca2+内流和超氧阴离子生成，而超氧化物清除显著抑制了METosis，且未改变NOX1/2蛋白水平，表明NOX的激活依赖于Ca2+。这些结果揭示了一条驱动LPS诱导METosis的SOCE-NOX-超氧化物轴，凸显其在肿瘤进展中的潜在作用。靶向SOCE有望破坏癌症发生和进展中巨噬细胞介导的炎症环路。
查看英文原文 English abstract
Macrophage extracellular traps (METs) contribute to chronic inflammation in the tumor microenvironment, promoting cancer progression via necroptosis-induced extracellular matrix degradation. This study investigates the role of store-operated calcium entry (SOCE) in calcium-induced MET formation, a process that amplifies pro-tumorigenic inflammatory responses in macrophages. Mouse macrophage RAW264.7 and human monocyte U937 cells were stimulated with lipopolysaccharide (LPS) or phorbol myristate acetate (PMA), with or without pretreatment using Ca 2+ chelator BAPTA-AM, mechanical Ca 2+ channel inhibitor GdCl 3 , SOCE inhibitors SKF96365 or YM58483, or superoxide anion scavenger thymoquinone (TQ). METotic cells were identified via immunofluorescence staining for Lamin B after digitonin permeabilization. LPS and PMA triggered SOCE-mediated Ca 2+ influx, leading to elevated cytoplasmic Ca 2+ levels that activated NADPH oxidase (NOX) to produce superoxide anions, culminating in MET formation. Pharmacological inhibition of SOCE attenuated Ca 2+ influx and superoxide anion, whereas superoxide scavenging significantly inhibited METosis without altering NOX1/2 protein levels, indicating Ca 2+ -dependent NOX activation. These results reveal a SOCE-NOX-superoxide axis driving LPS-induced METosis, highlighting its potential in tumor progression. Targeting SOCE could disrupt macrophage-mediated inflammatory circuits in cancer initiation and progression
利益披露 Disclosure
N. Nguyen, None..
W. Qiu, None.