Screening of differentially expressed proteins related to metastasis in hepatocellular carcinoma based on iTRAQ

Song Linlan; Yang Min; Jiang Xiaoyan; Chen Yu; Tang Hui; Yan Liang; Liao Yong

doi:10.13406/j.cnki.cyxb.001026

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Screening of differentially expressed proteins related to metastasis in hepatocellular carcinoma based on iTRAQ

更新时间：2024-01-17

- Screening of differentially expressed proteins related to metastasis in hepatocellular carcinoma based on iTRAQ
- Journal of Chongqing Medical University Vol. 41, Issue 10, Pages: 983-989(2016)
- 作者机构：
  
  重庆医科大学病毒性肝炎研究所教育部感染性疾病分子生物学重点实验室重庆医科大学附属第二医院感染科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001026
  CLC： R735.7
- Published：2016，
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[1]宋琳岚,杨敏,蒋小燕,陈瑜,汤慧,颜亮,廖勇.基于iTRAQ技术筛选肝癌转移相关差异蛋白[J].重庆医科大学学报,2016,41(10):983-989.

Song Linlan, Yang Min, Jiang Xiaoyan, et al. Screening of differentially expressed proteins related to metastasis in hepatocellular carcinoma based on iTRAQ. [J]. Journal of Chongqing Medical University 41(10):983-989(2016)
[1]宋琳岚,杨敏,蒋小燕,陈瑜,汤慧,颜亮,廖勇.基于iTRAQ技术筛选肝癌转移相关差异蛋白[J].重庆医科大学学报,2016,41(10):983-989. DOI： 10.13406/j.cnki.cyxb.001026.

Song Linlan, Yang Min, Jiang Xiaoyan, et al. Screening of differentially expressed proteins related to metastasis in hepatocellular carcinoma based on iTRAQ. [J]. Journal of Chongqing Medical University 41(10):983-989(2016) DOI： 10.13406/j.cnki.cyxb.001026.

摘要

目的:通过对不同转移潜能的肝癌细胞系的差异蛋白组学研究

以筛选鉴定出与肝癌转移相关的差异表达蛋白。方法:3株具有相同遗传背景、不同转移潜能的肝癌细胞系MHCC97-L、MHCC97-H及HCCLM3作为研究材料

经过收集胞质蛋白

浓缩、酶解、脱盐并用同位素标记相对和绝对定量技术（isobaric tags for relative and absolute quantitation

iTRAQ）对其进行标记

应用二维液相色谱串联质谱技术对肽段进行鉴定

得到的结果与数据库进行比对

对蛋白进行注释。基于Web Gestalt数据库对差异表达的蛋白进行GO（gene ontology）功能分析和通路分析以筛选与肝癌转移相关的目标蛋白。并用Western blot对3个关键蛋白（PLEC、SHMT2、LDHA）进行验证。结果:经过iTRAQ标记并质谱检测分析

共发现鉴定1 170个蛋白

其中在L/H（MHCC97-L/MHCC9-H）组鉴定发现47个差异表达蛋白

在M/H（HCCLM3/MHCC97-H）组鉴定发现65个差异表达蛋白。生物信息学分析显示L/H和M/H组差异表达的蛋白主要参与酶催化和细胞间粘附及细胞代谢的过程。应用Western blot在蛋白水平上的验证

PLEC在高转移细胞株MHCC97-H和HCCLM3中表达上调

SHMT2在HCCLM3中表达上调

LDHA在MHCC97-H中表达上调。结论:在本研究中应用iTRAQ标记高通量的蛋白质组学技术对3株不同转移潜能的肝癌细胞系的差异表达蛋白进行检测和鉴定分析

发现PLEC和SHMT2蛋白是两种新的与肝癌转移可能相关的蛋白。

Abstract

Objective:To investigate proteomics of hepatocellular carcinoma(HCC)cells with different metastatic potentials by isobaric tags for relative and absolute quantitation(i TRAQ);to screen and identify proteins related to metastasis

and to provide new molecular targets for metastasis and treatment. Methods:Cytoplasmic proteins

extracted from three cell lines with similar genetic background and different metastatic potentials were concentrated

digested

desalted and labeled with i TRAQ. Differentially expressed proteins were identified by 2D-LC-MS /MS

then the proteins were annotated by comparing with known protein database. Target proteins related with metastasis were singled out by gene ontology analysis and related pathway analysis using Web Gestalt databases. These proteins were then validated by Western blot. Results:Using i TRAQ labeling for quantification and detection by LC-MS/MS

1 170 proteins were identified. Forty-seven proteins were differential expressed in L/H(MHCC97-L/MHCC9-H)groups

and 65 proteins in M/H(HCCLM3/MHCC97-H)groups. Bioinformatics analysis revealed that differential expression proteins in L/H and M/H groups were mainly related to the catalytic activity of some enzymes and cells adhesion

and play important role in the cellular metabolic process. At translational level

PLEC was up-regulated in MHCC97-H and HCCLM-3

SHMT2 was up-regulated in HCCLM3

LDHA was up-regulated in MHCC97-H

thus the results of Western blot were consistent with those of LC-MS/MS. Conclusion:PLEC and SHMT2 are validated to be novel candidate HCC metastasis-associated proteins based on the analysis of this study.

关键词

肝细胞性肝癌蛋白组学同位素标记相对和绝对定量技术转移

Keywords

hepatocellular carcinomaproteomicsisobaric tags for relative and absolute quantitationmetastasis

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