Comparative proteome analysis of human esophageal cancer and adjacent normal tissues

Document Type: Original Article


1 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Cancer Research Center, Department of Radiation oncology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Pharmacology and Toxicology, School of Pharmacy, Shiraz University of Medical Sciences, Iran

6 Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

7 Nanotechnology Research Center, Department of Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad Iran


Objective(s): Ranking as the sixth commonest cancer, esophageal squamous cell carcinoma (ESCC) represents one of the leading causes of cancer death worldwide. One of the main reasons for the low survival of patients with esophageal cancer is its late diagnosis.
Materials and Methods: We used proteomics approach to analyze ESCC tissues with the aim of a better understanding of the malignant mechanism and searching candidate protein biomarkers for early diagnosis of esophageal cancer. The differential protein expression between cancerous               and normal esophageal tissues was investigated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Then proteins were identified by matrix-assisted laser desorption/ ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and MASCOT web based search engine.
Results:We reported 4 differentially expressed proteins involved in the pathological process of esophageal cancer, such as annexinA1 (ANXA1), peroxiredoxin-2 (PRDX2), transgelin (TAGLN) andactin-aortic smooth muscle (ACTA2).
Conclusion: In this report we have introduced new potential biomarker (ACTA2). Moreover, our data confirmed some already known markers for EC in our region.


1. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden ofcancer in 2008. Int J Cancer 2010; 127:2893-2917.

2. Qi YJ, Chao WX, Chiu JF. An overview of esophageal squamous cell carcinoma proteomics. J Proteomics 2012;8;75:3129-3137.

3. Rice TW, Rusch VW, Apperson-Hansen C, Allen MS, Chen LQ, Hunter JG, et al. World wide esophageal cancer collaboration. Dis Esophagus 2009; 22:1-8.

4. Cho HJ, Baek KE, Park SM, Kim IK, Choi YL, Cho HJ, et al. RhoGDI2 expression is associated with tumor growth and malignant progression of gastric cancer. Clin Cancer Res 2009; 15:2612-2619.

5. Hongsachart P, Huang-Liu R, Sinchaikul S, Pan FM, Phutrakul S, Chuang YM, et al. Glycoproteomic analysis of WGA-bound glycoprotein biomarkers in sera from patients with lungadenocarcinoma.Electrophoresis 2009; 30:1206-1220.

6. Schulz DM, Bollner C, Thomas G, Atkinson M, Esposito I, Hofler H, et al. Identification of differentially expressed proteins in triple-negative breast carcinomas using DIGE and mass spectrometry.J Proteome Res. 2009;8:3430-3438.

7. Mathivanan S, Lim JW, Tauro BJ, Ji H, Moritz RL, Simpson RJ. Proteomics analysis of A33immunoaffinity-purified exosomes released from the human colon tumor cell line LIM1215 reveals atissue-specific protein signature. Mol Cell Proteomics 2010;9:197-208.

8. Ono M, Matsubara J, Honda K, Sakuma T, Hashiguchi T, Nose H, et al. Prolyl 4-hydroxylation ofalpha-fibrinogen: a novel protein modification revealed by plasma proteomics. J Biol Chem 2009; 284:29041-29049.

9. Vellaichamy A, Sreekumar A, Strahler JR, Rajendiran T, Yu J, Varambally S, et al. Proteomic interrogation of androgen action in prostate cancer cells reveals roles of aminoacyl tRNA synthetases. PLoS One 2009; 18;4:e7075

10. Uemura N, Kondo T. Current advances in esophageal cancer proteomics. Biochim Biophys Acta 2014: 1854:687-695.

11. Moghanibashi M, Zare M, Jazii FR. Proteomics and Esophageal Cancer. esophageal cancer–cell and molecular biology, biomarkers, nutrition and treatment 2012:83-85.

12. Uemura N, Kondo T. Current advances in esophageal cancer proteomics. Biochimicaet BiophysicaActa (BBA)-Proteins and Proteomics 2014: 1854:687-695.

13. Moghanibashi M, Jazii FR, Soheili Z-S, Zare M, Karkhane A, Parivar K, et al. Proteomics of a newesophageal cancer cell line established from Persian patient. Gene 2012; 500:124-133.

14. Jazii FR, Najafi Z, Malekzadeh R, Conrads TP, Ziaee AA, Abnet C, et al. Identification of squamouscell carcinoma associated proteins by proteomics and loss of beta tropomyosin expression in esophagealcancer. World journal of gastroenterology: WJG. 2006; 12:7104-7012.

15. Mi H, Lazareva-Ulitsky B, Loo R, Kejariwal A, Vandergriff J, Rabkin S, et al. The PANTHERdatabase of protein families, subfamilies,  functions and pathways. Nucleic Acids Res 2005; 33: 284-288.

16. Flower RJ, Rothwell NJ. Lipocortin-1: cellular mechanisms and clinical relevance. Trends Pharmacol Sci 1994;15:71-76.

17. Perretti M, Gavins FN. Annexin 1: an endogenous anti-inflammatory protein. News Physiol Sci 2003;18:60-64.

18. Mussunoor S, Murray GI. The role of annexins in tumour development and progression. J Pathol 2008; 216:131-140.

19. Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev. 2002; 82:331-3371.

20. Bastian BC. Annexins in cancer and autoimmune diseases. Cell Mol Life Sci 1997; 53:554-556.

21. Paweletz CP, Ornstein DK, Roth MJ, Bichsel VE, Gillespie JW, Calvert VS, et al. Loss of annexin 1correlates with early onset of tumorigenesis in esophageal and prostate carcinoma. Cancer Res.2000;60:6293-6297.

22. Hippo Y, Yashiro M, Ishii M, Taniguchi H, Tsutsumi S, Hirakawa K, et al. Differential geneexpression profiles of scirrhous gastric cancer cells with high metastatic potential to peritoneum orlymph nodes. Cancer Res 2001; 61:8890-8895.

23. Garcia Pedrero JM, Fernandez MP, Morgan RO, Herrero Zapatero A, Gonzalez MV, Suarez Nieto C, et al. Annexin A1 down-regulation in head and neck cancer is associated with epithelial differentiationstatus. Am J Pathol 2004; 164:73-79.

24. Schwartz-Albiez R, Koretz K, Moller P, Wirl G. Differential expression of annexins I and II innormal and malignant human mammary epithelial cells. Differentiation 1993; 52:229-237.

25. Shen D, Chang HR, Chen Z, He J, Lonsberry V, Elshimali Y, et al. Loss of annexin A1 expression inhuman breast cancer detected by multiple high-throughput analyses. Biochem Biophys Res Commun 2005; 326:218-227.

26. Alldridge LC, Harris HJ, Plevin R, Hannon R, Bryant CE. The annexin protein lipocortin 1 regulatesthe MAPK/ERK pathway. J Biol Chem 1999; 274:37620-37628.

27. Alldridge LC, Bryant CE. Annexin 1 regulates cell proliferation by disruption of cell morphologyand inhibition of cyclin D1 expression through sustained activation of the ERK1/2 MAPK signal. Exp CellRes 2003; 290:93-107.

28. Shen D, Nooraie F, Elshimali Y, Lonsberry V, He J, Bose S, et al. Decreased expression of annexinA1 is correlated with breast cancer development and progression as determined by a tissue microarrayanalysis. Hum Pathol. 2006; 37:1583-1591.

29. Xia SH, Hu LP, Hu H, Ying WT, Xu X, Cai Y, et al. Three isoforms of annexin I are preferentiallyexpressed in normal esophageal epithelia but down-regulated in esophageal squamous cell carcinomas.Oncogene 2002; 21:6641-6648.

30. Gao Y, Chen Y, Xu D, Wang J, Yu G. Differential expression of ANXA1 in benign humangastrointestinal tissues and cancers. BMC cancer 2014; 14:520-562.

31. Wang KL, Wu TT, Resetkova E, Wang H, Correa AM, Hofstetter WL, et al. Expression of annexinA1 in esophageal and esophagogastric junction adeno-carcinomas: association with poor outcome. Clin Cancer Res 2006; 12:4598-4604.

32. Assinder SJ, Stanton JA, Prasad PD. Transgelin: an action-binding protein and tumour suppressor. ‎Int JBiochem Cell Biol 2009; 41:482-486.

33. Prasad PD, Stanton JA, Assinder SJ. Expression of the actin-associated protein transgelin (SM22) is decreased in prostate cancer. Cell Tissue Res 2010; 339:337-347.

34. Shields JM, Rogers-Graham K, Der CJ. Loss of transgelin in breast and colon tumors and in RIE-1cells by Ras deregulation of gene expression through Raf-independent pathways. J Biol Chem2002; 277:9790-9799.

35. Moghanibashi M, Jazii FR, Soheili ZS, Zare M, Karkhane A, Parivar K, et al. Proteomics of a newesophageal cancer cell line established from Persian patient. Gene. 2012;500:124-133.

36. Qi Y, Chiu JF, Wang L, Kwong DL, He QY. Comparative proteomic analysis of esophagealsquamous cell carcinoma.Proteomics 2005; 5:2960-2971.

37. Pawar H, Kashyap MK, Sahasrabuddhe NA, Renuse S, Harsha HC, Kumar P, et al. Quantitative tissue proteomics of esophageal squamous cell carcinoma for novel biomarker discovery. Cancer Biol Ther. 2011; 12:510-522.

38. Dvorakova M, Nenutil R, Bouchal P. Transgelins, cytoskeletal proteins implicated in differentaspects of cancer development. Expert Rev Proteomics. 2014;11:149-165.

39. Qi YJ, He QY, Ma YF, Du YW, Liu GC, Li YJ, et al. Proteomic identification of malignanttransformation-related proteins in esophageal squamous cell carcinoma. J Cell Biochem 2008; 104:1625-1635.

40. Rhee SG, Chae HZ, Kim K. Peroxiredoxins: a historical overview and speculative preview of novelmechanisms and emerging concepts in cell signaling. ‎Free Radic Biol Med 2005; 38:1543-1552.

41. Ishii T, Warabi E, Yanagawa T. Novel roles of peroxiredoxins in inflammation, cancer and innateimmunity. J Clin Biochem Nutr 2012; 50:91-105.

42. Carta F, Demuro PP, Zanini C, Santona A, Castiglia D, D'Atri S, et al. Analysis of candidate genesthrough a proteomics-based approach in primary cell lines from malignant melanomas and theirmetastases. Melanoma. Res 2005; 15:235-244.

43. Memon AA, Chang JW, Oh BR, Yoo YJ. Identification of differentially expressed proteins duringhuman urinary bladder cancer progression. Cancer Detect Prev 2005; 29:249-255.

44. Noh DY, Ahn SJ, Lee RA, Kim SW, Park IA, Chae HZ. Overexpression of peroxiredoxin in humanbreast cancer. Anticancer res 2001; 21:2085-2090.

45. Lee KW, Lee DJ, Lee JY, Kang DH, Kwon J, Kang SW. Peroxiredoxin II Restrains DNA Damageinduced Death in Cancer Cells by Positively Regulating JNK-dependent DNA Repair. J Biol Chem 2011; 286:8394-8404.

46. Du XL, Hu H, Lin DC, Xia SH, Shen XM, Zhang Y, et al. Proteomic profiling of proteins dysregultedin Chinese esophageal squamous cell carcinoma. J Mol Med 2007; 85:863-875.

47. Fu L, Qin YR, Xie D, Chow HY, Ngai SM, Kwong DL, et al. Identification of alpha-actinin 4 and 67kDalaminin receptor as stage-specific markers in esophageal cancer via proteomic approaches. Cancer 2007; 110:2672-2681.

48. Liu Z, Feng JG, Tuersun A, Liu T, Liu H, Liu Q, et al. Proteomic identification of differentially expressed proteins in esophageal cancer in three ethnic groups in Xinjiang. Mol Biol Rep. 2011; 38:3261-3269.

49. Ren HZ, Pan GQ, Wang JS, Wen JF, Wang KS, Luo GQ,      et al. Reduced stratify in expression canserve as an independent prognostic factor for poor survival in patients with esophageal squamous cell carcinoma. Dig Dis Sci 2010; 55:2552-2560.

50. Nishimori T, Tomonaga T, Matsushita K, Oh-Ishi M, Kodera Y, Maeda T, et al. Proteomic analysisof primary esophageal squamous cell carcinoma reveals downregulation of a cell adhesion protein,periplakin. Proteomics 2006; 6:1011-1018.

51. Hatakeyama H, Kondo T, Fujii K, Nakanishi Y, Kato H, Fukuda S, et al. Protein clusters associatedwith carcinogenesis, histological differentiation and nodal metastasis in esophageal cancer. Proteomics 2006; 6:6300-6316.

52. Zhang H-Z, Jin G-F, Shen H-B. Epidemiologic differences in esophageal cancer between Asian and Western populations.Chin J Cancer 2012; 31:281–286.

53. Li Y, Qin X, Cui J, Dai Z, Kang X, Yue H, et al. Proteome analysis of aflatoxin B1-inducedhepatocarcinogenesis in tree shrew (Tupaiabelangerichinensis) and functional identifi-cation ofcandidate protein peroxiredoxin II. Proteomics 2008; 8:1490-1501.

54. Lindberg U, Karlsson R, Lassing I, Schutt CE, Hoglund AS. The microfilament system and malignancy. Semin Cancer Biol 2008; 18:2-11.19.

55. Lee HW, Park YM, Lee SJ, Cho HJ, Kim DH, Lee JI, et al. Alpha-smooth muscle actin (ACTA2) isrequired for metastatic potential of human lung adenocarcinoma. Clin Cancer Res 2013; 19:5879-5889.

56. Sinn M, Denkert C, Striefler JK, Pelzer U, Stieler JM, Bahra M, et al. alpha-Smooth muscle actinexpression and desmoplastic stromal reaction in pancreatic cancer. BJC 2014; 111:1917-1923.