凝胶电泳-毛细管液相色谱-质谱联用技术分析大肠癌肿瘤与癌旁组织的差异蛋白质组

引用本文: 刘卓, 李洪军, 李晓鸥, 杨照微, 赵丽纯, 黄丽红, 郭宏华, 王海, 赵晴, 何成彦. 凝胶电泳-毛细管液相色谱-质谱联用技术分析大肠癌肿瘤与癌旁组织的差异蛋白质组[J]. 分析化学, 2017, 45(3): 322-330. doi: 10.11895/j.issn.0253-3820.160851 shu

Citation: LIU Zhuo, LI Hong-Jun, LI Xiao-Ou, YANG Zhao-Wei, ZHAO Li-Chun, HUANG Li-Hong, GUO Hong-Hua, WANG Hai, ZHAO Qing, HE Cheng-Yan. Differential Proteomics Analysis of Tumor Tissues and Their Adjacent Normal Tissues from Patients with Colorectal Cancer by Gel Electrophoresis-Capillary Liquid Chromatography-Mass Spectrometry-based Approach[J]. Chinese Journal of Analytical Chemistry, 2017, 45(3): 322-330. doi: 10.11895/j.issn.0253-3820.160851 shu

凝胶电泳-毛细管液相色谱-质谱联用技术分析大肠癌肿瘤与癌旁组织的差异蛋白质组

刘卓 ^1, ,
李洪军 ^1, ,
李晓鸥 ^2, ,
杨照微 ^1, ,
赵丽纯 ^3, ,
黄丽红 ^1, ,
郭宏华 ^1, ,
王海 ^1, ,
赵晴 ^1, ,
何成彦 ^1,

1.
吉林大学中日联谊医院, 长春 130033;
2.
吉林省肿瘤医院, 长春 130021;
3.
吉林大学药学院, 长春 130023
基金项目:
本文系国家自然科学基金(Nos.81572082，81472030)、吉林省科技厅项目(Nos.2011713，20150414015GH，20150101153JC，20140311092YY，2014041005GH)资助

摘要: 通过凝胶电泳将大肠癌患者的手术标本组织(肿瘤与癌旁)中的总蛋白按照分子量分成不同的组分，经胶内水解后，利用毛细管液相色谱-质谱联用技术对各个组分的多肽混合物进行蛋白质组学分析，共鉴定出29种差异表达的蛋白质，其中肿瘤组织中表达上调的蛋白质19种，如Fibrinogen beta chain，Vimentin，Fibrinogen gamma chain，Histone H2A type 1-B/E，Isoform 1 of Periostin，Fibrinogen alpha chain，Histone H3.1，Alpha-enolase，Protein disulfide-isomerase A3等；在肿瘤组织中表达下调的有10种蛋白质，如Sarcomeric tropomyosin kappa，Transgelin，Heat shock protein beta-1，Talin-1，Isoform 2 of Vinculin，Isoform 1 of Filamin-C等。通过蛋白质印迹实验对其中的踝蛋白1(Talin-1)和烯醇化酶(Alpha-enolase)的蛋白质组学鉴定结果进行了验证。生物信息学分析表明在大肠癌组织中表达上调的蛋白多具有胞外分泌的属性，提示这些蛋白很有可能会渗透到机体内的循环系统中，从而在患者血液或者腹水中能够被检测到；而在大肠癌组织中表达下调的蛋白则多为细胞骨架、胞外基质纤维等的组成成分，这些蛋白的下调常与肿瘤细胞向周围组织的侵袭和转移有密切关系。本研究为大肠癌的早期诊断、大肠癌初筛等方面的研究等提供了基础数据，对进一步深入研究大肠癌发病机制具有重要意义。

关键词:

English

Differential Proteomics Analysis of Tumor Tissues and Their Adjacent Normal Tissues from Patients with Colorectal Cancer by Gel Electrophoresis-Capillary Liquid Chromatography-Mass Spectrometry-based Approach

1.
China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2.
Tumor Hospital of Jilin Province, Changchun 130021, China;
3.
College of Pharmacology, Jilin University, Changchun 130023, China
Received Date: 22 November 2016
Revised Date: 13 January 2017
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 81572082, 81472030) and Jilin Province Science and Technology Deparement, China (Nos. 2011713, 20150414015GH, 20150101153JC, 20140311092YY, 2014041005GH)

Abstract: The whole protein extracts from tissue specimens (tumor and noncancerous) from patients with colorectal cancer were divided into several fractions with different molecular weight by gel electrophoresis, which were then subject to in-gel digestion and proteomics analysis by capillary liquid chromatography-mass spectrometry. A total of 29 differentially expressed proteins were identified, among which 19 were up-regulated proteins in tumor tissues, such as Fibrinogen beta chain, Vimentin, Fibrinogen gamma chain, Histone H2A type 1-B/E, Isoform 1 of Periostin, Fibrinogen alpha chain, Histone H3.1, Alpha-enolase, Protein disulfide-isomerase A3, and so on. Ten proteins were found to be down-regulated, including Sarcomeric tropomyosin kappa, Transgelin, Heat shock protein beta-1, Talin-1, Isoform 2 of Vinculin, Isoform 1 of Filamin-C, and so on. The differential expressions of Talin-1 and Alpha-enolase were selected to be verified by Western blot analysis. Bioinformatics analysis results indicated that most of the up-regulated proteins in colorectal cancer tissues had the feature of protein exocytosis, suggesting that these proteins were likely to penetrate into the circulatory system in the body, which could be detected in blood or ascites. On the other hand, most of the down-regulated proteins belonged to the cytoskeleton and extracellular matrix fibers, which were closely related to the invasion and metastasis of tumor cells into their surrounding tissues. This study provided basic data for the early diagnosis of colorectal cancer, colorectal cancer screening and other aspects, which had important significance to further study the pathogenesis of colorectal cancer.

Key words:

Gel electrophoresis-capillary liquid chromatography-mass spectrometry
/ Colorectal cancer
/ Proteome
/ Talin 1
/ Enolase 1

1. [1]
  LAI Yue-Xing,JING Da-Dao. China Cancer,2007,5:323-326 赖跃兴, 靖大道.中国肿瘤,2007,5:323-326
2. [2]
  Mármol I, Sánchez-de-Diego C, Pradilla Dieste A, Cerrada E, Rodriguez Yoldi M J. Int. J. Mol. Sci., 2017,18(1):197
3. [3]
  Guina T, Biasi F, Calfapietra S, Nano M, Poli G. Ann. N. Y. Acad. Sci., 2015,1340:95-103
4. [4]
  Giusti L, Iacconi P, Da Valle Y, Ciregia F, Ventroni T, Donadio E, Giannaccini G, Chiarugi M, Torregrossa L, Proietti A, Basolo F, Lucacchini A. Mol. Biosyst., 2012,8(4):1088-1099
5. [5]
  Hammoudi A, Song F, Reed K R, Jenkins R E, Meniel V S, Watson A J, Pritchard D M, Clarke A R, Jenkins J R. Biochem. Biophys. Res. Commun., 2013,440(3):364-370
6. [6]
  Yamamoto T, Kudo M, Peng W X, Takata H, Takakura H, Teduka K, Fujii T, Mitamura K, Taga A, Uchida E, Naito Z. Tumour Biol., 2016,37(10):13595-13606
7. [7]
  Yu K H, Snyder M. Mol. Cell. Proteomics, 2016,15(8):2525-2536
8. [8]
  WANG Zi-Jian, GAO Xiang, YANG Jing-Bo, SUN Wan-Chun, WU Dong-Lin, PENG Qi-Sheng, LIU Ning. Chinese J. Anal. Chem., 2016, 44(10):1521-1527 王子健, 高祥, 杨静波, 孙万春, 吴东林, 彭其胜, 刘宁.分析化学, 2016, 44(10):1521-1527
9. [9]
  BAI Hai-Xin, LIU Xiao-Hua, YANG Fan, YANG Xiu-Rong, WANG Er-Kang. Chem. J. Chinese Universities,2010,31(7):1314-1321 白海鑫, 刘小花, 杨帆, 杨秀荣, 汪尔康.高等学校化学学报,2010,31(7):1314-1321
10. [10]
  LIN Lin, LUO Shu-Sheng, WANG Lin-Jue, YANG Jie, SHEN Hai-Nan, TIAN Rui-Jun. Chinese J. Anal. Chem., 2015,43(10):1479-1489 林琳, 罗树生, 王灵珏, 杨杰, 沈海南, 田瑞军.分析化学,2015,43(10):1479-1489
11. [11]
  Broeckx V, Boonen K, Pringels L, Sagaert X, Prenen H, Landuyt B, Schoofs L, Maes E. Mol. Biosyst., 2016,12(2):553-565
12. [12]
  Cutillas P R, Crnogorac-Jurcevic T. Methods Mol. Biol., 2016,1381:201-209
13. [13]
  WU Peng, HE Fu-Chu, JIANG Ying. Prog. Biochem. Biophys., 2013, 40(3):281-292 武鹏, 贺福初, 姜颖.生物化学与生物物理进展,2013, 40(3):281-292
14. [14]
  Washburn M P, Wolters D, Yates J R 3rd. Nat. Biotechnol., 2001,19(3):242-247
15. [15]
  Haining A W, Lieberthal T J, Del Río Hernández A. FASEB J., 2016,30(6):2073-2085
16. [16]
  Fang K P, Zhang J L, Ren Y H, Qian Y B. Asian Pac. J. Cancer Prev., 2014,15(6):2655-2661
17. [17]
  SUN Li, ZHAO Hai-Feng, GUO Hong-Hua, YANG Guang-Yuan, HE Cheng-Yan, SHI Qing-Hong, WEN Xue, WANG Xiao-Ting, ZHAO Li-Chun. Chinese J. Anal. Chem., 2012,40(10):1521-1527 孙莉, 赵海丰, 郭宏华, 杨光远, 何成彦, 师庆红, 文雪, 王小婷, 赵丽纯.分析化学, 2012,10(40):1500-1506
18. [18]
  Shi Q N, Gao S, Song L N, Zhao Y M, Li X O, Wu J L, He C Y, Li H J, Zhao H F. Proteome Sci., 2015,13(1):1

扫一扫看文章

计量

PDF下载量: 2
文章访问数: 481
HTML全文浏览量: 42

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

凝胶电泳-毛细管液相色谱-质谱联用技术分析大肠癌肿瘤与癌旁组织的差异蛋白质组

English

Differential Proteomics Analysis of Tumor Tissues and Their Adjacent Normal Tissues from Patients with Colorectal Cancer by Gel Electrophoresis-Capillary Liquid Chromatography-Mass Spectrometry-based Approach

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

凝胶电泳-毛细管液相色谱-质谱联用技术分析大肠癌肿瘤与癌旁组织的差异蛋白质组

English

Differential Proteomics Analysis of Tumor Tissues and Their Adjacent Normal Tissues from Patients with Colorectal Cancer by Gel Electrophoresis-Capillary Liquid Chromatography-Mass Spectrometry-based Approach

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs