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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (3): 22-26.doi: 10.6040/j.issn.1671-7554.0.2014.800

• 基础医学 • 上一篇    下一篇

HPV永生化细胞系与SiHa细胞系蛋白质表达差异的质谱分析

刘亚飞, 张奇舒, 袁首道, 陈健行   

  1. 山东大学癌症研究中心, 山东 济南 250012
  • 收稿日期:2014-11-08 修回日期:2015-01-23 出版日期:2015-03-10 发布日期:2015-03-10
  • 通讯作者: 陈健行, E-mail:jjc12345.chen@gmail.com E-mail:jjc12345.chen@gmail.com

Mass spectrometry analysis of differentially expressed proteins between HPV immortalized cell line and SiHa cell line

LIU Yafei, ZHANG Qishu, YUAN Shoudao, Jason J. Chen   

  1. Cancer Research Centre, Shandong University, Jinan 250012, Shandong, China
  • Received:2014-11-08 Revised:2015-01-23 Online:2015-03-10 Published:2015-03-10

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摘要： 目的 分析人乳头瘤病毒(HPV)永生化细胞系与SiHa细胞系之间的蛋白质表达差异, 探讨从HPV感染到宫颈癌发生间的变化。方法 高效液相色谱(LC)与质谱(MS)联用对HPV永生化细胞系与SiHa细胞系的蛋白质进行鉴定, 然后通过蛋白质组学和生物信息学分析两种细胞系之间的蛋白表达,及一种新型翻译后修饰-琥珀酰化修饰的差异。结果 在两种细胞系中, 质谱鉴定到的蛋白总数为10 691个,琥珀酰化修饰位点共6 342个;非标定量分析共发现572种差异表达蛋白。HPV永生化细胞系中高表达的273种蛋白多集中在疾病的相关通路, 而宫颈癌细胞系中高表达的299种蛋白多集中在代谢通路和核糖体通路。琥珀酰化修饰无差异, 但发现一个新的琥珀酰化修饰位点——血清和糖皮质激素激酶1(SGK1) K41琥珀酰化。结论 与HPV永生化细胞系相比, SiHa细胞系代谢能力及翻译蛋白能力更强;而HPV永生化细胞中高表达蛋白则大多与疾病相关, 且更易发生恶化。

关键词: 人乳头瘤病毒, 蛋白质组学, 质谱, 宫颈肿瘤

Abstract: Objective To explore the mechanism by which human papilloma virus (HPV) induces cervical cancer and to identify proteins differentially expressed between HPV immortalized cells and SiHa cells. Methods Differentially expressed proteins between HPV immortalized cells and SiHa cells were identified by high performance liquid chromatography and mass spectrometry, and analyzed by label-free quantity software and bioinformatics approaches. Results A total of 10 691 proteins were identified and 6 342 succinylation sites were found; label-free quantification analyses indicated that 572 proteins were differentially expressed between the two types of cells; bioinfomatic analysis showed that 273 proteins in HPV immortalized cells were mostly associated with disease pathway, while 299 proteins in cervical cancer cells were mostly involved in metabolic and protein-produced pathway. Although succinylation modifications had no differences, a new succinylation site at SGK1 K41 was found. Conclusion Compared with HPV immortalized cells, SiHa cells demonstrate higher level of metabolism and stronger protein-synthesis, while most up-regulated proteins in HPV immortalized cell are associated with particular diseases.

Key words: Proteomics, Human papilloma virus, Mass spectrometry, Cervical neoplasm

中图分类号: 

  • R737.33
[1] Aggarwal P. Cervical cancer:Can it be prevented?[J]. World J Clin Oncol, 2014, 5(4):775-780.
[2] Shigehara K, Sasagawa T, Namiki M. Human papillomavirus infection and pathogenesis in urothelial cells:a mini-review[J]. J Infect Chemother, 2014, 20(12):741-747.
[3] Zheng ZM, Baker CC. Papillomavirus genome structure, expression, and post-transcriptional regulation [J]. Front Biosci, 2006, 11(16):2286-2302.
[4] 李华, 高国兰. HPV与宫颈癌的研究进展[J]. 实用癌症杂志, 2007, 4(20):420-422.
[5] 梁东霞, 张彦娜. 宫颈癌与HPV关系的研究进展[J]. 实用癌症杂志, 2010, 2(25):202-205.
[6] Mazurek S. Pyruvate kinase type M2:A key regulator of the metabolic budget system in tumor cells [J]. Int J Biochem Cell Biol, 2011, 43(7):969-980.
[7] Zhou Z, Kim J, Insolera R, et al. Rho GTPase regulation of α-synuclein and VMAT2:implications for pathogenesis of Parkinson's disease [J]. Mol Cell Neurosci, 2011, 48(1):29-37.
[8] Fan X, Liu Y, Heilman SA, et al. Human papillomavirus E7 induces rereplication in response to DNA damage [J]. J Virol, 2013, 87(2):1200-1210.
[9] Karim S, Mirza Z, Kamal MA, et al. Gene expression analysis approach to establish possible links between parkinson's disease, cancer and cardiovascular diseases [J]. CNS Neurol Disord Targets, 2014, 13(8):1334-1345.
[10] Delaney LM, Ho N, Morrison J, et al. Dichloroacetate affects proliferation but not survival of human colorectal cancer cells [J]. Apoptosis, 2014, doi.10.1007/s10495-014-1046-4.
[11] Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect:the metabolic requirements of cell proliferation [J]. Science, 2009, 324(5930):1029-1033.
[12] Ma Z, Vosseller K. Cancer Metabolism and Elevated O-GlcNAc in Oncogenic Signaling[J]. J Biol Chem, 2014, 289(50):34457-34465.
[13] Warner SL, Carpenter KJ, Bearss DJ. Activators of PKM2 in cancer metabolism [J]. Future Med Chem, 2014, 6(10):1167-1178.
[14] Xie Z, Dai J, Dai L, et al. Lysine succinylation and lysine malonylation in histones [J]. Mol Cell Proteomics, 2012, 11(5):100-107.
[15] Zheng J. Energy metabolism of cancer:Glycolysis versus oxidative phosphorylation[J]. Oncol Lett, 2012, 4(6):1151-1157.
[16] Du J, Zhou Y, Su X, et al. Sirt5 is a NAD-dependent protein lysine demalonylase and desuccinylase[J]. Science, 2011, 334(6057):806-809.
[17] 詹成, 时雨, 马军, 等. 丙酮酸激酶M2型(PKM2)入核机制和核内作用的研究进展[J]. 复旦学报:医学版, 2014, 41(1):133-137. ZHAN Cheng, SHI Yu, MA Jun, et al. A review of the mechanism of pyruvate kinase M2's nuclear translocation and its function in nucleus[J]. Fudan University Journal of Medical Sciences, 2014, 41(1):133-137.
[18] Li P, Pan F, Hao Y, et al. SGK1 is regulated by metabolic-related factors in 3T3-L1 adipocytes and overexpressed in the adipose tissue of subjects with obesity and diabetes[J]. Diabetes Res Clin Pract, 2013, 102(1):35-42.
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