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Zhiyu Zhang et al.Quantitative proteomic analysis of glycosylated proteins enriched from urine samples with magnetic ConA nanoparticles identifies potential biomarkers for small cell lung,Journal of Pharmaceutical and Biomedical Analysis,Available online 31 August 2021, 114352

ABSTRACT

Lung cancer has high morbidity and mortality and small cell lung cancer (SCLC) is a highly invasive malignant tumor with a very unfavorable survival rate. Early diagnosis and treatment can result in better prognosis for the SCLC patients but current diagnostic methods are either invasive or incapable for large-scale screen. Therefore, discovering biomarkers for early diagnosis of SCLC is of importance. In this work, we covalently coupled Concanavalin A (ConA) to functionalized magnetic nanoparticles to obtain magnetic ConA-nanoparticles (ConA-NPs) for the enrichment of glycosylated proteins. We then purified glycosylated proteins in 36 urine samples from 9 healthy controls, 9 SCLC patients, 9 lung adenocarcinoma (LUAD) patients, and 9 squamous cell lung carcinoma (LUSC) patients. The purified glycosylated proteins were digested and analyzed by LC-MS/MS for identification and quantification. Among the 398 identified proteins, 20, 15, and 1 glycosylated protein(s), respectively, were upregulated in the urine of SCLC, LUAD, and LUSC patients. Immunoblotting experiments further demonstrated that cathepsin C and transferrin were significantly upregulated in the ConA-NP purified urine of SCLC patients. This work suggests that glycosylated cathepsin C and transferrin might be able to serve as potential biomarkers for the noninvasive diagnosis of SCLC patients.

肺癌具有較高的發(fā)病率和死亡率，小細(xì)胞肺癌（SCLC）是一種侵襲性很強(qiáng)的惡性腫瘤，生存率非常低。早期診斷和治療可以改善小細(xì)胞肺癌患者的預(yù)后，但目前的診斷方法要么是侵入性的，要么不能進(jìn)行大規(guī)模篩查。因此，發(fā)現(xiàn)SCLC早期診斷的生物標(biāo)志物具有重要意義。在這項(xiàng)工作中，我們將刀豆球蛋白A（ConA）與功能化磁性納米顆粒(PuriMag G-COOH)共價(jià)偶聯(lián)，以獲得用于富集糖基化蛋白質(zhì)的磁性ConA納米顆粒（ConA NPs）。然后我們從9名健康對(duì)照者、9名小細(xì)胞肺癌患者、9名肺腺癌（LUAD）患者和9名肺鱗狀細(xì)胞癌（LUSC）患者的36份尿液樣本中純化糖基化蛋白。純化的糖基化蛋白經(jīng)LC-MS/MS消化和分析，用于鑒定和定量。在398種已鑒定的蛋白中，SCLC、LUAD和LUSC患者的尿液中分別有20、15和1種糖基化蛋白上調(diào)。免疫印跡實(shí)驗(yàn)進(jìn)一步證明，小細(xì)胞肺癌患者的ConA NP純化尿液中組織蛋白酶C和轉(zhuǎn)鐵蛋白顯著上調(diào)。這項(xiàng)研究表明，糖基化組織蛋白酶C和轉(zhuǎn)鐵蛋白可以作為小細(xì)胞肺癌患者無創(chuàng)性診斷的潛在生物標(biāo)志物。

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