Extracellular vesicles derived from ovarian cancer cell lines discriminated by biochemical and Fourier transform infrared spectroscopy approaches†

IF 3.3 3区 化学 Q2 CHEMISTRY, ANALYTICAL Analyst Pub Date : 2025-02-28 DOI:10.1039/D5AN00024F
Lefkothea Pantazi, Valérie Untereiner, Paolo Rosales, Romain Rivet, Sandra Audonnet, Isabelle Proult, Laurent Ramont, Ganesh D. Sockalingum and Stéphane Brézillon
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Abstract

Ovarian cancer is the most lethal cancer among gynaecological malignancies. Due to the lack of early symptoms and screening tools, patients are diagnosed in advanced stages. Cancer invasion and metastasis through the extracellular matrix (ECM) are enhanced by tumour cell Extracellular Vesicles (EV). The aim of this study was to characterise the EVs derived from two ovarian cancer cell lines (ES2 and SKOV3) using biochemical and vibrational spectroscopic approaches. EVs were prepared by ultracentrifugation and characterised by Nanoparticle Tracking Analysis. Their surface proteins were assessed by MACSPlex EV kit for human exosomes. The presence of MMP14 and integrin subunits was evaluated in EVs and cell protein extracts by Western immunoblotting. Both EVs and cells were measured by Fourier transform infrared spectroscopy (FTIR) and data were analysed by hierarchical cluster analysis (HCA). Spectral differences were observed in the lipids and polysaccharides regions both between the SKOV3 and ES2 cells and their corresponding EVs, which allowed a good delineation by HCA. The differences in the biochemical data were confirmed by similar and specific features exhibited in their respective infrared spectral signatures. ES2 EVs exhibited an enrichment in MMP14 in agreement with the aggressiveness of this ovarian cancer metastatic cell line.

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用生化和傅里叶变换红外光谱方法鉴别卵巢癌细胞系的细胞外囊泡
卵巢癌是妇科恶性肿瘤中最致命的癌症。由于缺乏早期症状和筛查工具,患者在晚期被诊断出来。肿瘤细胞胞外囊泡(EV)增强了肿瘤通过细胞外基质(ECM)的侵袭和转移。本研究的目的是利用生化和振动光谱方法表征来自两种卵巢癌细胞系(ES2和SKOV3)的ev。采用超离心法制备了电动汽车,并采用纳米颗粒跟踪分析对其进行了表征。用人外泌体MACSPlex EV试剂盒检测其表面蛋白。Western免疫印迹法检测EVs和细胞蛋白提取物中MMP14和整合素亚基的存在。采用傅里叶变换红外光谱(FTIR)对电动汽车和电池进行测量,并用层次聚类分析(HCA)对数据进行分析。在SKOV3和ES2细胞及其相应的ev之间,脂质区和多糖区光谱存在差异,这使得HCA可以很好地描绘。在各自的红外光谱特征中表现出相似和特定的特征,证实了生化数据的差异。ES2 ev表现出MMP14的富集,这与该卵巢癌转移细胞系的侵袭性一致。
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来源期刊
Analyst
Analyst 化学-分析化学
CiteScore
7.80
自引率
4.80%
发文量
636
审稿时长
1.9 months
期刊介绍: "Analyst" journal is the home of premier fundamental discoveries, inventions and applications in the analytical and bioanalytical sciences.
期刊最新文献
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