Department of Clinical Laboratory, Harbin Medical University Cancer Hospital, Harbin, 150081, Heilongjiang, China.
Department of Pathology, The First Affiliated Hospital of Jiamusi University, Jiamusi, 154003, Heilongjiang, China.
Cell Mol Life Sci. 2024 Jan 10;81(1):19. doi: 10.1007/s00018-023-05051-9.
Cardiovascular disorders are commonly prevalent in cancer patients, yet the mechanistic link between them remains poorly understood. Because neutrophil extracellular traps (NETs) have implications not just in cardiovascular diseases (CVD), but also in breast cancer (BC), it was hypothesized to contribute to CVD in the context of oncogenesis. We established a mouse model using nude mice to simulate liver metastasis of triple-negative BC (TNBC) through the injection of MDA-MB-231 cells. Multiple imaging and analysis techniques were employed to assess the cardiac function and structure, including echocardiography, HE staining, Masson staining, and transmission electron microscopy (TEM). MDA-MB-231 cells underwent treatment with a CaSR inhibitor, CaSR agonist, and NF-κB channel blocker. The phosphorylation of NF-κB channel protein p65 and the expression and secretion of IL-8 were assessed using qRT-PCR, Western Blot, and ELISA, respectively. In addition, MDA-MB-231 cells were co-cultured with polymorphonuclear neutrophils (PMN) under varying conditions. The co-localization of PMN extracellular myeloperoxidase (MPO) and DNA were observed by cellular immunofluorescence staining to identify the formation of NETs. Then, the cardiomyocytes were co-cultured with the above medium that contains NETs or not, respectively; the effects of NETs on cardiomyocytes apoptosis were perceived by flow cytometry. The ultrastructural changes of myocardial cells were perceived by TEM, and ELISA detected the levels of myocardial enzyme (LDH, MDA and SOD). Overall, according to our research, CaSR has been found to have a regulatory role in IL-8 secretion in MDA-MB-231 cells, as well as in the formation of NETs by PMN cells. These findings suggest CaSR-mediated stimulation in PMN can lead to increased NETs formation and subsequently to cytotoxicity in cardiomyocytes, which potentially via activation of the NF-κB signaling cascade of BC cell.
心血管疾病在癌症患者中普遍存在，但它们之间的机制联系仍知之甚少。由于中性粒细胞胞外诱捕网（NETs）不仅与心血管疾病（CVD）有关，也与乳腺癌（BC）有关，因此有人假设它在肿瘤发生过程中会导致 CVD。我们通过注射 MDA-MB-231 细胞，建立了一个使用裸鼠模拟三阴性乳腺癌（TNBC）肝转移的小鼠模型。我们采用多种成像和分析技术来评估心脏功能和结构，包括超声心动图、HE 染色、Masson 染色和透射电子显微镜（TEM）。MDA-MB-231 细胞接受钙敏感受体（CaSR）抑制剂、激动剂和 NF-κB 通道阻滞剂的处理。通过 qRT-PCR、Western blot 和 ELISA 分别评估 NF-κB 通道蛋白 p65 的磷酸化以及 IL-8 的表达和分泌。此外，MDA-MB-231 细胞在不同条件下与多形核粒细胞（PMN）共培养。通过细胞免疫荧光染色观察 PMN 细胞外髓过氧化物酶（MPO）和 DNA 的共定位，以鉴定 NETs 的形成。然后，将上述含有或不含有 NETs 的培养基与心肌细胞共培养，通过流式细胞术观察 NETs 对心肌细胞凋亡的影响。TEM 观察心肌细胞的超微结构变化，ELISA 检测心肌酶（LDH、MDA 和 SOD）水平。总的来说，根据我们的研究，CaSR 被发现对 MDA-MB-231 细胞中 IL-8 的分泌以及 PMN 细胞中 NETs 的形成具有调节作用。这些发现表明，PMN 中 CaSR 介导的刺激可导致 NETs 形成增加，并随后导致心肌细胞的细胞毒性，这可能是通过 BC 细胞 NF-κB 信号级联的激活。
