3种不同的HPV型别宫颈癌细胞株分泌蛋白非标记定量技术分析

doi:10.3969/j.issn.1004-616x.2018.05.005

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (5): 354-358,364.doi: 10.3969/j.issn.1004-616x.2018.05.005

3种不同的HPV型别宫颈癌细胞株分泌蛋白非标记定量技术分析

郝轶^1,3, 袁建林², 肖悦¹, 李卉³, 郭霞^1,3

1. 南方医科大学深圳医院, 广东深圳 518100;
2. 新疆医科大学附属肿瘤医院, 新疆乌鲁木齐 830011;
3. 新疆医科大学地方病分子生物学实验室, 新疆乌鲁木齐 830011

收稿日期:2018-05-21 修回日期:2018-09-13 出版日期:2018-09-30 发布日期:2018-09-30
通讯作者: 郭霞,E-mail:myshow0504@163.com E-mail:myshow0504@163.com
作者简介:郝轶,E-mail:haoyi0320@126.com。
基金资助:
新疆维吾尔自治区自然科学基金（2015211C004）

Label-free quantitative proteomics analysis of secretory proteins from three different HPV-types of cervical cancer cell lines

HAO Yi^1,3, YUAN Jianlin², XIAO Yue¹, LI Hui³, GUO Xia^1,3

1. Shenzhen Hospital, Southern Medical University, Shenzhen 518100, Guangdong;
2. Affiliated Cancer Hospital, Xinjiang Medical University, Urumqi 830011;
3. Xinjiang Key Laboratory of Molecular Biology and Endemic Diseases, Xinjiang Medical University, Urumqi 830011, Xinjiang, China

Received:2018-05-21 Revised:2018-09-13 Online:2018-09-30 Published:2018-09-30

摘要/Abstract

摘要： 目的：分析3种不同HPV型别的宫颈癌细胞株Siha、Caski和C33的分泌蛋白表达及种类，寻找其共性分泌蛋白标志物，为临床早期发现及针对性干预提供依据。方法：采用3种不同宫颈癌SiHa、Caski和C33细胞株的分泌蛋白，运用LC-MS/MS和Maxquant的非标记技术，筛选出宫颈癌分泌蛋白的种类及分类。结果：共鉴定肽段数21 609个，蛋白数2 895个。其中3种宫颈癌细胞株共性蛋白共729种，占29.2%；特异性蛋白Siha细胞株含128种，Caski细胞株含143种，C33细胞株含859种。对于结果中729种共性蛋白进行分析，有以下4类较为集中的类型：热休克蛋白类（HSPs）共14种；异质性核糖核蛋白类（HNRP）共8种；层黏连蛋白类（LN）共4种；阿尔多酮还原酶1类（AKR1）共2种。其中，溶酶体信号通路所占比例最大（39种）。结论：SiHa、Caski和C33细胞株的分泌蛋白中共有729种共性蛋白，其中4类集中的蛋白类型均先后在研究中证实对宫颈癌的进程和发展有影响，并且可能与溶酶体信号通路有关。可为筛选高危型HPV感染的宫颈癌患者的血清标志物提供依据。

关键词: 宫颈癌, 宫颈癌细胞株, 分泌蛋白, 非标记定量技术分析

Abstract: OBJECTIVE:To analyze secretory proteins from three different kinds of cervical cancer cell lines (Siha, Caski and C33) and to identify specific proteins as biomarkers for earlier detection and for treatment. METHODS:Label-free quantitative proteomics technology processes were performed successively:protein cleavage, protein quantification and proteolysis. Then,the secretory proteins were analyzed and were picked out by LC-MS/MS and unlabeled Maxquant analyses. RESULTS:21 609 kinds of peptides and 2 895 different kinds of proteins were found. Meanwhile,collective proteins made up 29.2% of total with 729 different types. The numbers of proteins for Siha,Caski and C33 were 128,143 and 859,respectively. Among the collective proteins,there were 4 main protein species:heat shock proteins (HSPs)(14 proteins), heterogeneous nuclear ribonucleoprotein (HNRP)(8 proteins), laminin (LN)(4 proteins) and Aldo-keto reductase family 1(AKR1) (2 proteins). In addition,the lysosomal signaling pathway accounted for the largest proportion (39 proteins). CONCLUSION:There were 729 kinds of collective proteins among the three cervical cancer cell lines which belonged to the HPV16 type. Among the collective proteins,4 main protein species were concluded which have already been confirmed to play a significant role in the cervical cancer process. They may also belong to the lysosomal signaling pathway,The results may provide certain research directions for tumor molecular markers in the future.

Key words: cervical cancer, cell lines of cervical cancer, secretory protein, label-free analysis

中图分类号:

R730.3

郝轶, 袁建林, 肖悦, 李卉, 郭霞. 3种不同的HPV型别宫颈癌细胞株分泌蛋白非标记定量技术分析[J]. 癌变·畸变·突变, 2018, 30(5): 354-358,364.

HAO Yi, YUAN Jianlin, XIAO Yue, LI Hui, GUO Xia. Label-free quantitative proteomics analysis of secretory proteins from three different HPV-types of cervical cancer cell lines[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(5): 354-358,364.

参考文献

[1] 潘林. ZEBI在宫颈组织中的表达和临床意义[J]. 现代肿瘤杂志,2016,24(10):1613-1617.
[2] MBEUNKUI F, METGE B J, SHEVDE L A, et al. Identification of differentially secreted biomarkers using LC MS/MS in isogenic cell lines representing a progression of breast cancer[J]. J Proteome Res,2007,6(8):2993-3002.
[3] 张永谦,来成俊,李琰,等. OFFGEL和LC-MS联用分析SH-SY5Y细胞分泌蛋白质组[J]. 北京理工大学学报,2011, 31(10):1252-1255.
[4] COX J, MANN M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification[J]. Nat Biotechnol,2008,26(12):1367-1372.
[5] COX J, NEUHAUSER N, MICHALSKI A, et al. Andromeda:a peptide search engine integrated into the MaxQuant environment[J]. J Proteome Res, 2011, 10(4):1794-1805.
[6] NO J H,KIM M K,JEON Y T,et al. Human papillomavirus vaccine:widening the scope for cancer prevention[J]. Mol Carcinog,2011,50(4):244-253.
[7] BZHALAVA D, GUAN P, FRANCESCHI S, et al. A systematic review of the prevalence of mucosal and cutaneous human papillomavirus types[J]. Virology, 2013, 445:224-231.
[8] 刘会兰,徐慧,青碧兰. HPV病毒16型感染的影响因素研究[J]. 国际病毒学杂志,2016,23(1):60-63.
[9] WAANDERS L F, CHWALEK K, MONETTI M, et al. Quantitative proteomic analysis of single pancreatic islets[J]. Proc Natl Acad Sci USA,2009,106(45):18902-18907.
[10] LUBER C A,COX J,LAUTERBACH H et al. Quantitative proteomics reveals subset-specific viral recognition in dendritic cells[J]. Immunity,2010,26,32(2):279-289.
[11] KARAGIANNIS G S, PAVLOU M P, DIAMANDIS E P. Cancer secretomics reveal pathophysiological pathways in cancer molecular oncology[J]. Mol Oncol,2010,4:496-510.
[12] 李贤煜,应万涛,钱小红. 癌细胞分泌蛋白质组富集与鉴定策略研究进展[J]. 生物技术通讯,2013,24(3):430-436.
[13] 李明,邓芳,王保龙. 分泌蛋白质组学在肿瘤标志物研究中的进展[J]. 临床输血与检验,2013,15(4):407-409.
[14] 党军强,罗健康,任摇涛. 分泌蛋白质组学在肿瘤标志物研究中的现状及进展[J]. 现代肿瘤医学,2011,19(12):2576-2579.
[15] KEIKO Y,AKIHIRO M,KAZUYUKI Y,et al. Heat shock protein 70 is involved in malignant behaviorsand chemosensitivities to cisplatin in cervical squamous cell carcinoma cells[J]. J Obstet Gynaecol Res, 2014, 40(5):1188-1196.
[16] XUE DU,YONGMEI LI,XU JING,et al. Effect of a heat shock protein 90 specific inhibitor on the proliferation and apoptosis induced by VEGFC in cervical cancer cells[J]. Exp Ther Med,2014,8:1559-1564.
[17] MANOJ G, DEEPIKA K, SHIKHA S, et al. Germ cell specific heat shock protein 70-2 is expressed in cervical carcinoma and is involved in the growth, migration, and invasion of cervical cells[J]. Cancer,2010:3785-3795.
[18] 赵恩锋,鲍嫘,梁龙,李冬萍. 热休克蛋白在宫颈癌和癌前病变中的表达[J]. 中国应用生理学杂志,2006,22(2):250-253.
[19] SONIYA D, NAOKO K, JACOB G, et al. Heterogeneous nuclear ribonucleoprotein C proteins interact with the human papillomavirus type 20(HPV16) early 3'-untranslated region and alleviate suppression of HPV16 late L1 mRNA splicing[J]. J Biol Chem,2015,290(21):13354-13371.
[20] AJAY M, SATYENDRA C T, DESOUZA L V, et al. Heterogeneous ribonucleoprotein K is a marker of oral leukoplakia and correlates with poor prognosis of squamous cell carcinoma[J]. Int J Cancer,2009,125:1398-1406.
[21] HIROYASU KASHIMA, REN-CHIN WU, YIHONG WANG, et al. Laminin C1 expression by uterine carcinoma cells is associated with tumor progression[J]. Gynecol Oncol, 2015,139(2):338-344.
[22] YOSHITAKE H, TAKAHASHI M, ISHIKAWA H, et al.Aldo-keto reductase family 1, member B10 in uterine carcinomas:a potential risk factor of recurrence after surgical therapy in cervical cancer[J]. Int J Gynecol Cancer,2007, 17:1300-1306.
[23] MARIO S,BHAVIN S,MICHAEL H,et al. Entry of human papillomavirus type 16 by actin-dependent,clathrin-and lipid Raft-independent endocytosis[J]. PLoS Pathog,2012,8(4):e1002657.
[24] JASON M,ARI H. Virus entry by micropinocytosis[J]. Nat Cell Biol,2009,11(5):510-520.
[25] SPODEN G,FREITAG K,HUSMANN M,et al. Clathrin and caveolin-independent entry of human papillomavirus type 16 involvement of tetraspanin-enriched microdomains (TEMs)[J]. PLoS One,2008,3:3313.

3种不同的HPV型别宫颈癌细胞株分泌蛋白非标记定量技术分析

Label-free quantitative proteomics analysis of secretory proteins from three different HPV-types of cervical cancer cell lines

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