Clinical Proteomics | |
Aberrant Mucin5B expression in lung adenocarcinomas detected by iTRAQ labeling quantitative proteomics and immunohistochemistry | |
Qing Kay Li3  Hui Zhang1  Frederic Askin1  Edward Gabrielson1  MingHui Ao1  Xiangchun Wang1  Yan Li2  | |
[1] Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA;Current address: Institute of Biophysics, Room 9214, 15 Datun Road, Room 9214, Chanyang, Beijing 100101, P. R. China;Department of Pathology, The Johns Hopkins Bayview Medical Center, 4940 Eastern Ave., AA Building, Room 154, Baltimore, MD 21224, USA | |
关键词: Immunohistochemistry; Quantitative proteomics and iTRAQ labeling; Expression of mucin5B; Lung adenocarcinoma; Non-small cell lung cancer; | |
Others : 1026335 DOI : 10.1186/1559-0275-10-15 |
|
received in 2013-04-02, accepted in 2013-09-17, 发布年份 2013 | |
【 摘 要 】
Background
Lung cancer is the number one cause of cancer-related deaths in the United States and worldwide. The complex protein changes and/or signature of protein expression in lung cancer, particularly in non-small cell lung cancer (NSCLC) has not been well defined. Although several studies have investigated the protein profile in lung cancers, the knowledge is far from complete. Among early studies, mucin5B (MUC5B) has been suggested to play an important role in the tumor progression. MUC5B is the major gel-forming mucin in the airway. In this study, we investigated the overall protein profile and MUC5B expression in lung adenocarcinomas, the most common type of NSCLCs.
Methods
Lung adenocarcinoma tissue in formalin-fixed paraffin-embedded (FFPE) blocks was collected and microdissected. Peptides from 8 tumors and 8 tumor-matched normal lung tissue were extracted and labeled with 8-channel iTRAQ reagents. The labeled peptides were identified and quantified by LC-MS/MS using an LTQ Orbitrap Velos mass spectrometer. MUC5B expression identified by iTRAQ labeling was further validated using immunohistochemistry (IHC) on tumor tissue microarray (TMA).
Results
A total of 1288 peptides from 210 proteins were identified and quantified in tumor tissues. Twenty-two proteins showed a greater than 1.5-fold differences between tumor and tumor-matched normal lung tissues. Fifteen proteins, including MUC5B, showed significant changes in tumor tissues. The aberrant expression of MUC5B was further identified in 71.1% of lung adenocarcinomas in the TMA.
Discussions
A subset of tumor-associated proteins was differentially expressed in lung adenocarcinomas. The differential expression of MUC5B in lung adenocarcinomas suggests its role as a potential biomarker in the detection of adenocarcinomas.
【 授权许可】
2013 Li et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20140903113931127.pdf | 1998KB | download | |
Figure 5. | 105KB | Image | download |
Figure 4. | 34KB | Image | download |
Figure 3. | 35KB | Image | download |
Figure 2. | 71KB | Image | download |
Figure 1. | 76KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
【 参考文献 】
- [1]Siegel R, Naishadham D, Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 2012, 62:10-29.
- [2]Parkin M, Tyczynski JE, Boffetta P, Samet J, Shields P, Brambilla E, Caporaso N, Travis WD, Muller-Hermelink HK, Harris CC, Tumours of the lung (Eds): WHO classification of tumours. Pathology and genetics of tumours of the lung, pleura, thymus and heart. Lyon: IARC press; 2004:p9-124.
- [3]Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA: Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small cell lung cancer to gefitinib. N Engl J Med. 2004, 350:2129-2139.
- [4]Paez JG, Jänne PA, Lee JC, Tracy S, Greulich H, Gabriel S, Herman P, Kaye FJ, Lindeman N, Boggon TJ, Naoki K, Sasaki H, Fujii Y, Eck MJ, Sellers WR, Johnson BE, Meyerson M: EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science. 2004, 304:1497-1500.
- [5]Janku F, Garrido-Laguna I, Petruzelka LB, Stewart DJ, Kurzrock R: Novel therapeutic targets in non-small cell lung cancer. J Thorac Oncol 2011, 6:1601-1612. Review
- [6]Aberle DR, Adams AM, Berg CD, Black WC, Clapp JD, Fagerstrom RM, Gareen IF, Gatsonis C, Marcus PM, Sicks JD, National Lung Screening Trial Research Team: Reduced lung-cancer mortality with low-dose computed tomographic screening. N Engl J Med 2011, 365:395-409.
- [7]Beroukhim R, Mermel CH, Porter D, Wei G, Raychaudhuri S, Donovan J, Barretina J, Boehm JS, Dobson J, Urashima M, Mc Henry KT, Pinchback RM, Ligon AH, Cho YJ, Haery L, Greulich H, Reich M, Winckler W, Lawrence MS, Weir BA, Tanaka KE, Chiang DY, Bass AJ, Loo A, Hoffman C, Prensner J, Liefeld T, Gao Q, Yecies D, Signoretti S, et al.: The landscape of somatic copy-number alteration across human cancers. Nature. 2010, 463:899-905.
- [8]Anagnostou VK, Dimou AT, Botsis T, Killiam EJ, Gustavson MD, Homer RJ, Boffa D, Zolota V, Dougenis D, Tanoue L, Gettinger SN, Detterbeck FC, Syrigos KN, Bepler G, Rimm DL: Molecular classification of nonsmall cell lung cancer using a 4-protein quantitative assay. Cancer 2012, 118:1607-1618.
- [9]Chen G, Gharib TG, Wang H, Huang CC, Kuick R, Thomas DG, Shedden KA, Misek DE, Taylor JM, Giordano TJ, Kardia SL, Iannettoni MD, Yee J, Hogg PJ, Orringer MB, Hanash SM, Beer DG: Protein profiles associated with survival in lung adenocarcinoma. Proc Natl Acad Sci U S A. 2003, 100:13537-13542.
- [10]Nambu Y, Beer DG: Altered surface markers in lung cancer. Lack of cell-surface Fas/APO-1 expression in pulmonary adenocarcinoma may allow escape from immune surveillance. Methods Mol Med. 2003, 74:259-266.
- [11]Tian T, Hao J, Xu A, Luo C, Liu C, Huang L, et al.: Determination of metastasis-associated proteins in non-small cell lung cancer by comparative proteomic analysis. Cancer Sci 2007, 98:1265-1274.
- [12]Poletti V, Poletti G, Murer B, Saragoni L, Chilosi M: Bronchoalveolar lavage in malignancy. Semin Respir Crit Care Med. 2007, 28:534-545.
- [13]Yanagisawa K, Shyr Y, Xu BJ, Massion PP, Larsen PH, White BC, Roberts JR, Edgerton M, Gonzalez A, Nadaf S, Moore JH, Caprioli RM, Carbone DP: Proteomic patterns of tumour subsets in non-small-cell lung cancer. Lancet 2003, 362:433-439.
- [14]Kawamura T, Nomura M, Tojo H, Fujii K, Hamasaki H, Mikami S, Bando Y, Kato H, Nishimura T: Proteomic analysis of laser-microdissected paraffin-embedded tissues: (1) stage-related protein candidates upon non-metastatic lung adenocarcinoma. J Proteomics. 2010, 73:1089-1099.
- [15]Nishimura T, Nomura M, Tojo H, Hamasaki H, Fukuda T, Fujii K, Mikami S, Bando Y, Kato H: Proteomic analysis of laser-microdissected paraffin-embedded tissues: (2) MRM assay for stage-related proteins upon non-metastatic lung adenocarcinoma. J Proteomics. 2010, 73:1100-1110.
- [16]Chen G, Gharib TG, Huang CC, Thomas DG, Shedden KA, Taylor JM, Kardia SL, Misek DE, Giordano TJ, Iannettoni MD, Orringer MB, Hanash SM, Beer DG: Proteomic analysis of lung adenocarcinoma: identification of a highly expressed set of proteins in tumors. Clin Cancer Res. 2002, 8:2298-2305.
- [17]Li QK, Gabrielson E, Zhang H: Application of glycoproteomics in the discovery of biomarkers for lung cancer. Proteomics: Clinical App. 2012, 5–6:244-256.
- [18]Li QK, Gabrielson E, Askin F, Chan DW, Zhang H: Glycoproteomics using fluid based specimens in the discovery of lung cancer protein biomarkers. Promise and challenge. Proteomics Clinical App. 2013, 7:55-69.
- [19]Ross MC, Voynow JA: Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol Rev. 2006, 86:245-278.
- [20]Hollingsworth MA, Swanson BJ: Mucins in cancer: protection and control of the cell surface. Nature Rev Cancer. 2004, 4:45-60.
- [21]Dekker J, Rossen JW, Büller HA, Einerhand AW: The MUC family: an obituary. Trends Biochem Sci. 2002, 27:126-131.
- [22]Yu CJ, Yang PC, Shun CT, Lee YC, Kuo SH, Luh KT: Overexpression of MUC5 genes is associated with early post-operative metastasis in non-small-cell lung cancer. Int J Cancer. 1996, 69:457-465.
- [23]Awaya H, Takeshima Y, Yamasaki M, Inai K: Expression of MUC1, MUC2, MUC5AC, and MUC6 in atypical adenomatous hyperplasia, bronchioloalveolar carcinoma, adenocarcinoma with mixed subtypes, and mucinous bronchioloalveolar carcinoma of the lung. Am J Clin Pathol. 2004, 121:644-653.
- [24]Zhang H, Li XJ, Martin DB, Aebersold R: Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry. Nat Biotechnol. 2003, 21:660-666.
- [25]Tian Y, Tan AC, Sun X, Olson MT, Xie Z, Jinawath N, Chan DW, Shih IM, Zhang Z, Zhang H: Quantitative proteomic analysis of ovarian cancer cells identified mitochondrial proteins associated with Paclitaxel resistance. Proteomics Clin Appl 2009, 3:1288-1295.
- [26]Edge SB, Byrd DR, Compton CC, Fritz AG, Greene FL, Trotti A: AJCC Cancer Staging Manual. 7th edition. New York, NY: Springer; 2010.
- [27]Travis WD, Brambilla E, Noguchi M, et al.: International association for the study of lung cancer/American thoracic society/European respiratory society international multidisciplinary classification of lung adenocarcinoma. J Thorac Oncol. 2011, 6:244-285.
- [28]Silver SA, Askin FS: True papillary carcinoma of the lung: a distinct cliniclpathologic entity. Am J Surg Pathol 1997, 21:43-51.
- [29]Li QK, Shah P, Li Y, Aiyetan PO, Chen J, Yung RCW, Molena D, Gabrilson E, Askin F, Chan DW, Zhang H: Glycoproteomic analysis of bronchoalveolar lavage (BAL) fluid identifies tumor-associated glycoproteins from lung adenocarcinoma. J Proteome Res. 2013, 12:3638-3696.
- [30]Nakamori S, Ota DM, Cleary KR, Shirotani K, Irimura T: MUC1 mucin expression as a marker of progression and metastasis of human colorectal carcinoma. Gastroenterology. 1994, 106:353-361.
- [31]Lüttges J, Feyerabend B, Buchelt T, Pacena M, Klöppel G: The mucin profile of noninvasive and invasive mucinous cystic neoplasms of the pancreas. Am J Surg Pathol. 2002, 26:466-471.
- [32]Nitta T, Sugihara K, Tsuyama S, Murata F: Immunohistochemical study of MUC1 mucin in premalignant oral lesions and oral squamous cell carcinoma: association with disease progression, mode of invasion, and lymph node metastasis. Cancer. 2000, 88:245-254.
- [33]Takao S, Uchikura K, Yonezawa S, Shinchi H, Aikou T: Mucin core protein expression in extrahepatic bile duct carcinoma is associated with metastases to the liver and poor prognosis. Cancer. 1999, 86:1966-1975.
- [34]Komatsu M, Jepson S, Arango ME, Carothers Carraway CA, Carraway KL: Muc4/sialomucin complex, an intramembrane modulator of ErbB2/HER2/Neu, potentiates primary tumor growth and suppresses apoptosis in a xenotransplanted tumor. Oncogene. 2001, 20:461-470.
- [35]Perrais M, Pigny P, Copin MC, Aubert JP, Van Seuningen I: Induction of MUC2 and MUC5AC mucins by factors of the epidermal growth factor (EGF) family is mediated by EGF receptor/Ras/Raf/extracellular signal-regulated kinase cascade and Sp1. J Biol Chem. 2002, 277:32258-32267.
- [36]Copin MC, Buisine MP, Devisme L, Leroy X, Escande F, Gosselin B, Aubert JP, Porchet N: Normal respiratory mucosa, precursor lesions and lung carcinomas: differential expression of human mucin genes. Front Biosci 2001, 6:D1264-1275. Review
- [37]Copin MC, Buisine MP, Leteurtre E, Marquette CH, Porte H, Aubert JP, Gosselin B, Porchet N: Mucinous bronchioloalveolar carcinomas display a specific pattern of mucin gene expression among primary lung adenocarcinomas. Hum Pathol. 2001, 32:274-281.
- [38]MacDermed DM, Khodarev NN, Pitroda SP, Edwards DC, Pelizzari CA, Huang L, Kufe DW, Weichselbaum RR: MUC1-associated proliferation signature predicts outcomes in lung adenocarcinoma patients. BMC Med Genomics. 2010, 3:16. BioMed Central Full Text
- [39]Chen H, Suzuki M, Nakamura Y, Ohira M, Ando S, Iida T, Nakajima T, Nakagawara A, Kimura H: Aberrant methylation of FBN2 in human non-small cell lung cancer. Lung Cancer. 2005, 50:43-49.
- [40]Kukulj S, Jaganjac M, Boranic M, Krizanac S, Santic Z, Poljak-Blazi M: Altered iron metabolism, inflammation, transferrin receptors, and ferritin expression in non-small-cell lung cancer. Med Oncol. 2010, 27:268-277.
- [41]Oh JE, Krapfenbauer K, Fountoulakis M, Frischer T, Lubec G: Evidence for the existence of hypothetical proteins in human bronchial epithelial, fibroblast, amnion, lymphocyte, mesothelial and kidney cell lines. Amino Acids 2004, 26:8-18.
- [42]Konicek BW, Stephens JR, McNulty AM, Robichaud N, Peery RB, Dumstorf CA, Dowless MS, Iversen PW, Parsons S, Ellis KE, McCann DJ, Pelletier J, Furic L, Yingling JM, Stancato LF, Sonenberg N, Graff JR: Therapeutic inhibition of MAP kinase interacting kinase blocks eukaryotic initiation factor 4E phosphorylation and suppresses outgrowth of experimental lung metastases. Cancer Res. 2011, 71:1849-1857.
- [43]Bortner JD Jr, Das A, Umstead TM, Freeman WM, Somiari R, Aliaga C, et al.: Down-regulation of 14–3–3 isoforms and annexin A5 proteins in lung adenocarcinoma induced by the tobacco-specific nitrosamine NNK in the A/J mouse revealed by proteomic analysis. J Proteome Res 2009, 8:4050-4061.
- [44]Boersma HH, Kietselaer BL, Stolk LM, Bennaghmouch A, Hofstra L, Narula J, Heidendal GA, Reutelingsperger CP: Past, present, and future of annexin A5: from protein discovery to clinical applications. J Nucl Med 2005, 46:2035-2050. Review
- [45]Fritzsche FR, Dahl E, Dankof A, Burkhardt M, Pahl S, Petersen I, Dietel M, Kristiansen G: Expression of AGR2 in non small cell lung cancer. Histol Histopathol. 2007, 22:703-708.
- [46]Liu D, Rudland PS, Sibson DR, Platt-Higgins A, Barraclough R: Human homologue of cement gland protein, a novel metastasis inducer associated with breast carcinomas. Cancer Res. 2005, 65:3796-3805.
- [47]Xu N, Wang YS, Pan WB, Xiao B, Wen YJ, Chen XC, Chen LJ, Deng HX, You J, Kan B, Fu AF, Li D, Zhao X, Wei YQ: Human alpha-defensin-1 inhibits growth of human lung adenocarcinoma xenograft in nude mice. Mol Cancer Ther. 2008, 7:1588-1597.
- [48]Kikuchi T, Hassanein M, Amann JM, Liu Q, Slebos RJ, Rahman SM, Kaufman JM, Zhang X, Hoeksema MD, Harris BK, Li M, Shyr Y, Gonzalez AL, Zimmerman LJ, Liebler DC, Massion PP, Carbone DP: In-depth proteomic analysis of nonsmall cell lung cancer to discover molecular targets and candidate biomarkers. Mol Cell Proteomics. 2012, 11:916-32.