您的位置:山东大学 -> 科技期刊社 -> 《山东大学学报(医学版)》

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

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.
[1] 刘超,刁婷婷,张红吉,刘红伟,杨永锐,李晓非. 飞行时间质谱在耐药结核病检测中的应用价值[J]. 山东大学学报 (医学版), 2024, 62(4): 61-67.
[2] 梁中昊,庄向华,黄珊,韩晓琳,华梦羽,琚丽萍,陈诗鸿. 饮食诱导肥胖雌鼠棕色脂肪组织蛋白质组学分析[J]. 山东大学学报 (医学版), 2024, 62(2): 10-19.
[3] 石军,牛子捷,王军,马丽晶,奚春花,肖洋. 幼年型复发性呼吸道乳头状瘤病肺部播散的临床特点及麻醉要点[J]. 山东大学学报 (医学版), 2024, 62(12): 82-89.
[4] 于丹凤,曹娟,于世佳,杜鲁涛,侯新国. 电化学发光法与液相色谱-串联质谱法评估糖尿病患者维生素D缺乏程度的效能比较与界值确定[J]. 山东大学学报 (医学版), 2023, 61(7): 90-95.
[5] 刘萌,侯丛哲,麻焕玉,张震,赵新蕊,张萍,朱琳. β-雌二醇对宫颈癌Hela细胞增殖的影响[J]. 山东大学学报 (医学版), 2023, 61(2): 9-15.
[6] 李雁儒,李娟,李培龙,杜鲁涛,王传新. 胰腺癌不同进展期血清外泌体蛋白质组学分析[J]. 山东大学学报 (医学版), 2022, 60(10): 33-41.
[7] 关恒云,刘岚铮,姜慧钰,白爱英,刘辉,赵小冬,高尚,李娜,阮师漫. 利用时间质谱检出312例6月龄内健康婴儿鼻咽部微生态菌群的分布特征[J]. 山东大学学报 (医学版), 2022, 60(10): 49-56.
[8] 蔡春芳,易丹妮,郭芝亮,何耀娟. P16蛋白与TCT、HR-HPV的相关性及对不同子宫颈病变诊断的效能[J]. 山东大学学报 (医学版), 2022, 60(1): 40-47.
[9] 陈立晓,英信江,陈歆维,王菲,孙臻峰,董频. 下咽鳞癌蛋白质谱鉴定及预后靶分子筛选[J]. 山东大学学报 (医学版), 2021, 59(9): 140-147.
[10] 张华宇,殷思源,刘健,马嘉旭,宋茹,曹国起,王一兵. 氧糖剥夺条件下培养表皮干细胞的定量蛋白质组学分析[J]. 山东大学学报 (医学版), 2021, 59(4): 17-27.
[11] 刘冬科,王凤山. HILIC-MS/MS法鉴别肝素和硫酸乙酰肝素[J]. 山东大学学报 (医学版), 2021, 59(3): 41-47.
[12] 周慧梅,杨佳欣,曹冬焱,沈铿,向阳,吴鸣,潘凌亚,黄惠芳,郎景和. 早期宫颈癌保留生育功能手术的治疗效果及妊娠结局[J]. 山东大学学报 (医学版), 2018, 56(5): 18-22.
[13] 赵庆庆,尹格平. TNF-α 启动子5个位点的基因多态性与山东省汉族妇女宫颈癌遗传易感性的相关性[J]. 山东大学学报 (医学版), 2018, 56(2): 28-33.
[14] 张文浩,乔立君,郑静怡,陈健行,张魏芳. 人乳头瘤病毒早期癌蛋白E6/E7稳定表达细胞系的构建及其对Rab12表达的影响[J]. 山东大学学报 (医学版), 2018, 56(12): 1-6.
[15] 刘京康,杨建勇,孟丽华,姜洁. 血清miR-17-92簇在HPV阳性宫颈癌中的早期诊断价值[J]. 山东大学学报(医学版), 2017, 55(5): 86-90.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!