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基于iTRAQ的视网膜母细胞瘤蛋白质定量蛋白质组学分析及生物信息学研究

iTRAQ-based quantitative proteomic analysis and bioinformatics study of proteins in retinoblastoma.

作者信息

Cheng Yong, Meng Qingyu, Huang Lvzhen, Shi Xuan, Hou Jing, Li Xiaoxin, Liang Jianhong

机构信息

Ophthalmology Department, Peking University People's Hospital, Beijing 100044, P.R. China.

Key Laboratory of Vision Loss and Restoration, Ministry of Education, Beijing 100044, P.R. China.

出版信息

Oncol Lett. 2017 Dec;14(6):8084-8091. doi: 10.3892/ol.2017.7221. Epub 2017 Oct 19.

Abstract

The aim of the present study was to analyze proteins in the aqueous humor (AH) of patients' retinoblastoma (RB), and investigate their potential role in RB using the comparative proteomic technique of isobaric tags for relative and absolute quantitation (iTRAQ) coupled with offline two-dimensional liquid chromatography-tandem mass spectrometry. A total of 0.1 ml AH was collected from 10 children with RB (mean age, 3.8 years; range, 2-5 years) and patients with senile cataracts (mean age, 70.4 years; range, 65-79 years), which was used as the control. iTRAQ was used to analyze proteins in the AH of patients and controls. Proteins with a fold change of >1.20 or <0.83 were considered to be significantly differentially expressed (with corrected P<0.05). The identified proteins were subjected to subsequent gene ontology (GO) analysis using the DAVID database. A total of 83 proteins that were expressed differently between the controls and patients' AH samples were identified using iTRAQ analysis. Of these proteins, 44 were upregulated and 39 were downregulated. On the basis of biological processes in GO, the identified proteins were primarily involved in glycoprotein, amyloid acute-inflammatory and defensive responses. Among these proteins, pigment epithelium-derived factor serves a potential role in the treatment of RB, and stimulated by retinoic acid 6 may serve as a potential protein involved in RB development. To the best of our knowledge, the present study is the first to identify 83 proteins associated with RB using iTRAQ technology. The results of the present study will aid in furthering the understanding of RB and developing novel therapy targets in the future.

摘要

本研究的目的是分析视网膜母细胞瘤(RB)患者房水(AH)中的蛋白质,并使用相对和绝对定量的等压标签(iTRAQ)比较蛋白质组学技术结合离线二维液相色谱 - 串联质谱法,研究它们在RB中的潜在作用。从10例RB患儿(平均年龄3.8岁;范围2 - 5岁)和老年性白内障患者(平均年龄70.4岁;范围65 - 79岁)中收集共0.1 ml房水,后者用作对照。使用iTRAQ分析患者和对照的房水中的蛋白质。将倍数变化>1.20或<0.83的蛋白质视为差异有统计学意义(校正P<0.05)。使用DAVID数据库对鉴定出的蛋白质进行后续基因本体(GO)分析。使用iTRAQ分析鉴定出对照组和患者房水样本之间共83种表达不同的蛋白质。其中,44种上调,39种下调。基于GO中的生物学过程,鉴定出的蛋白质主要参与糖蛋白、淀粉样蛋白急性炎症和防御反应。在这些蛋白质中,色素上皮衍生因子在RB治疗中发挥潜在作用,而视黄酸刺激蛋白6可能是参与RB发生发展的潜在蛋白质。据我们所知,本研究首次使用iTRAQ技术鉴定出83种与RB相关的蛋白质。本研究结果将有助于进一步了解RB,并在未来开发新的治疗靶点。

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