Choezom Dolma, Plum Jan-Moritz, Karuna M Pradhipa, Danieli-Mackay Adi, Lenz Christof, Brockmeyer Phillipp, Gross Julia Christina
Department of Hematology and Oncology, University Medical Center Goettingen, 37075 Goettingen, Germany.
Department of Developmental Biochemistry, University Medical Center Goettingen, 37077 Goettingen, Germany.
Cells. 2025 Apr 4;14(7):547. doi: 10.3390/cells14070547.
Tumor-derived extracellular vesicles (EVs) play an important role in cancer progression. Neutral sphingomyelinases (nSMases) are lipid-modifying enzymes that modulate the secretion of EVs from cells. How nSMase activity and therefore ceramide generation affect the composition and functionality of secreted EVs is not fully understood. Here, we aimed to investigate the expression of nSMases 1 and 2 in prostate cancer (PCa) tissue and their role in EV composition and secretion for prostate cancer cell migration. Reduced nSMase 1 and 2 expression was found in prostate cancer and correlated with the age of the patient. When nSMase 2 was inhibited by GW4869 in PCa cells (PC3 and DU145), the EV secretome was significantly altered, while the number of EVs and the total protein content of released EVs were not significantly changed. Using proteomic analysis, we found that extracellular matrix proteins, such as SDC4 (Syndecan-4) and SRPX-2, were differentially secreted on EVs from GW4869-treated PC3 cells. In scratch wound migration assays, GW4869 significantly increased migration compared to control PC3 cells but not DU145 cells, while SDC4 knockdown significantly reduced the migration of PC3 cells. These and other nSMase-2-dependent secreted proteins are interesting candidates for understanding the role of stress-induced EVs in the progression of prostate cancer.
肿瘤衍生的细胞外囊泡（EVs）在癌症进展中起重要作用。中性鞘磷脂酶（nSMases）是脂质修饰酶，可调节细胞中EVs的分泌。nSMase活性以及由此产生的神经酰胺如何影响分泌的EVs的组成和功能尚不完全清楚。在这里，我们旨在研究nSMases 1和2在前列腺癌（PCa）组织中的表达及其在EV组成和分泌中对前列腺癌细胞迁移的作用。在前列腺癌中发现nSMase 1和2的表达降低，且与患者年龄相关。当用GW4869抑制PCa细胞（PC3和DU145）中的nSMase 2时，EV分泌组发生显著改变，而EVs的数量和释放的EVs的总蛋白含量没有显著变化。通过蛋白质组学分析，我们发现细胞外基质蛋白，如SDC4（Syndecan-4）和SRPX-2，在来自GW4869处理的PC3细胞的EVs上差异分泌。在划痕伤口迁移试验中，与对照PC3细胞相比，GW4869显著增加了迁移，但对DU145细胞没有影响，而SDC4敲低显著降低了PC3细胞的迁移。这些以及其他nSMase-2依赖性分泌蛋白是理解应激诱导的EVs在前列腺癌进展中的作用的有趣候选物。
