Ampudia-Mesias Elisabet, El-Hadad Samia, Cameron Charles Scott, Wöhrer Adelheid, Ströbel Thomas, Saydam Nurten, Saydam Okay
Department of Pediatrics, Division of Hematology and Oncology, Medical School, University of Minnesota, Minneapolis, MN 55454, USA.
Department of Reproductive Endocrinology, University Hospital Zurich, 8006 Zurich, Switzerland.
Cancers (Basel). 2022 Apr 14;14(8):1984. doi: 10.3390/cancers14081984.
Extracellular vesicles (EVs) may be used as a non-invasive screening platform to discover markers associated with early diagnosis, prognosis, and treatment response. Such an approach is invaluable for diseases such as glioblastoma, for which only a few non-invasive diagnostic or prognostic markers are available. We used mass spectrometry to analyze proteomics profiles of EVs derived from four glioblastoma cell lines and human primary astrocytes (HPAs) and found that SRPX is the only protein enriched in the majority of glioblastoma EVs that was absent in the HPA-derived EVs. Then, we evaluated the relationship between SRPX protein expression and tumor grade using immunohistochemical staining (IHC) and performed colony formation and viability assays to analyze the possible function of SRPX in glioblastoma. SRPX mRNA and protein expression were associated with tumor grade. Moreover, temozolomide (TMZ)-resistant tumor tissues showed highly positive SRPX staining, compared to all other tumor grades. Additionally, glioblastoma cells displayed enhanced SRPX gene expression when exposed to TMZ. Knockdown of SRPX gene expression via siRNA inhibited cell viability. Taken together, the results of this study suggest that SRPX can be used as a novel tumor marker for diagnostic and prognostic purposes and can also be a therapeutic target for glioblastomas.
细胞外囊泡(EVs)可用作非侵入性筛查平台,以发现与早期诊断、预后及治疗反应相关的标志物。对于胶质母细胞瘤这类仅有少数非侵入性诊断或预后标志物的疾病而言,这种方法具有重要价值。我们利用质谱分析法分析了源自四种胶质母细胞瘤细胞系和人原代星形胶质细胞(HPAs)的细胞外囊泡的蛋白质组学图谱,发现SRPX是多数胶质母细胞瘤细胞外囊泡中富集的唯一一种在源自人原代星形胶质细胞的细胞外囊泡中不存在的蛋白质。然后,我们使用免疫组织化学染色(IHC)评估了SRPX蛋白表达与肿瘤分级之间的关系,并进行了集落形成和活力测定,以分析SRPX在胶质母细胞瘤中的可能功能。SRPX mRNA和蛋白表达与肿瘤分级相关。此外,与所有其他肿瘤分级相比,耐替莫唑胺(TMZ)的肿瘤组织显示SRPX染色呈高度阳性。另外,胶质母细胞瘤细胞在暴露于替莫唑胺时显示出SRPX基因表达增强。通过小干扰RNA(siRNA)敲低SRPX基因表达可抑制细胞活力。综上所述,本研究结果表明SRPX可作为一种用于诊断和预后目的的新型肿瘤标志物,也可成为胶质母细胞瘤的治疗靶点。