Araştırma Makalesi
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Tümör Doku Heterojenitesinin 2DE Proteomik Yaklaşımıyla Gösterilmesi

Yıl 2022, , 301 - 314, 30.06.2022
https://doi.org/10.31832/smj.1066524

Öz

Amaç: Birçok hastalığın ve kanserlerin biyolojik mekanizmalarının incelenmesinde yaygın olarak kullanılan proteomik yaklaşımlar, hastalık sürecinin izlenmesinde, biyobelirteçlerin ve potansiyel terapötik hedeflerin tanımlanmasında kullanılagelmektedir. Proteomik alanında kullanılan biyolojik örnekler arasında serum gibi biyolojik sıvılar ve doku gibi katı örnekler sayılabilir. Bu bağlamda analiz edilen doku örneklerinin kalitesi, örneğin alındığı bölgenin doğruluğu ve hatta örneğin alınmasından sonra proteomik araştırma laboratuarına ulaşıncaya kadar geçen süre gibi pre-analitik süreçler çok önemlidir. Biz bu çalışmamızda, tümör dokusu içerisinde iki farklı bölgeden alınan örneklerin çözünür proteom profillerini karşılaştırmayı ve varsa tümör heterojenitesini göstermeyi amaçladık.
Gereç ve Yöntemler: Cushing sendromu tanısı ile opere edilen bir hastanın adrenokortikal tümör dokusunun iki farklı bölgesinden (adenoma ve adenoma içi) ve sağlıklı dokusundan elde edilen proteinler 2DE yöntemi ile ayrıştırılarak karşılaştırmalı analize tabi tutulmuştur. Örnekler arasında regülasyon düzeyinde farklılık görülen protein sayısı belirlenmiş ve jellerden kesilerek MALDI-TOF/TOF kütle spektrometresi aracılığı ile tanımlanmıştır.
Bulgular: Kontrol dokusuna göre adenom bölgesinde 17 protein, adenom içi bölgede ise 13 proteinin seviyelerinde regülasyon tespit edilmiştir. Bu proteinler çoğu enerji metabolizması, hücre iskeleti organizasyonu ve hücresel stres ile ilişkili proteinlerdir.
Sonuç: Bu çalışma tümör dokusu içerisinde örneğin alındığı bölgenin proteom profilini ne ölçüde etkilediğini göstermiştir. Tümör dokusundan iki farklı bölgeden aldığımız örneklerde daha çok enerji metabolizması ile ilişkili proteinler olmak üzere bazı proteinlerin ifadelerinde ciddi farklılıklar görülmüştür. Bu durum özellikle biyobelirteç arayışlarının olduğu çalışmalarda proteomik bulgularının yorumlanmasında dikkat edilmesi gerektiği ve proteomik çalışmalarında örnek sayısının mümkün olduğunca fazla tutulması gerektiğini vurgulamaktadır.

Destekleyen Kurum

Kocaeli Üniversitesi

Proje Numarası

KOÜ-BAP TSA-2020-1545

Teşekkür

Bu araştırma Kocaeli Üniversitesi Bilimsel Araştırma Birimi tarafından desteklenmiştir (Proje No: KOÜ-BAP TSA-2020-1545). Dokunun patolojik incelemesini yaptığı için Dr. Büşra Yaprak Bayrak’a teşekkür ederiz.

Kaynakça

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  • Referans30. Bansal V, El Asmar N, Selman WR, Arafah BM. Pitfalls in the diagnosis and management of Cushing's syndrome. Neurosurg Focus. Feb 2015;38(2):E4. doi:10.3171/2014.11.FOCUS14704
  • Referans31. Clayton RN, Raskauskiene D, Reulen RC, Jones PW. Mortality and morbidity in Cushing's disease over 50 years in Stoke-on-Trent, UK: audit and meta-analysis of literature. J Clin Endocrinol Metab. Mar 2011;96(3):632-42. doi:10.1210/jc.2010-1942
  • Referans32. Duan K, Hernandez KG, Mete O. Clinicopathological correlates of adrenal Cushing's syndrome. Postgrad Med J. Jun 2015;91(1076):331-42. doi:10.1136/postgradmedj-2014-202612rep
  • Referans33. Kjellin H, Johansson H, Hoog A, Lehtio J, Jakobsson PJ, Kjellman M. Differentially expressed proteins in malignant and benign adrenocortical tumors. PLoS One. 2014;9(2):e87951. doi:10.1371/journal.pone.0087951
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  • Referans35. Kotlowska A, Malinski E, Sworczak K, Kumirska J, Stepnowski P. The urinary steroid profile in patients diagnosed with adrenal incidentaloma. Clin Biochem. Apr 2009;42(6):448-54. doi:10.1016/j.clinbiochem.2008.12.027
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The Demonstration of Tumor Heterogeneity using 2DE Proteomic Approach

Yıl 2022, , 301 - 314, 30.06.2022
https://doi.org/10.31832/smj.1066524

Öz

Objective: Proteomic approaches, have been used in monitoring the disease processes and identification of biomarkers. Biological samples used in the field of proteomics include biological fluids and solid samples. In this context, pre-analytical processes e.g., tissue sample quality, sampling site accuracy and sample transfer time to the site of proteomic laboratory are very important. In this study, the proteome profiles of samples taken from two different regions within a tumor tissue were compared to show tumor heterogeneity.
Material and Methods: Proteins obtained from two different regions of adrenocortical tumor tissue (adenoma and intra-adenoma) and healthy tissue of a patient who was operated on with the diagnosis of Cushing's syndrome were separated by 2DE and subjected to comparative analysis. The number of differentially regulated proteins among the samples was determined and identified by MALDI-TOF/TOF mass spectrometry.
Results: Seventeen and thirteen proteins were differentially regulated in the adenoma and intra adenoma regions compared to the control tissue, respectively. These proteins mostly associated with energy metabolism, cytoskeletal organization, and cellular stress.
Conclusion: This study showed to what extent the sample area in the tumor tissue affects the proteome profile. The samples taken from two different regions of a same tumor tissue displayed serious differences in the expressions of some proteins, mostly those associated with energy metabolism. This highlights the need to be careful in interpreting proteomic findings, especially in studies where biomarkers are sought. To minimize sample variation, as many samples as possible should be studied in proteomic studies.

Proje Numarası

KOÜ-BAP TSA-2020-1545

Kaynakça

  • Referans1. Anderson NG, Anderson NL. Twenty years of two-dimensional electrophoresis: past, present and future. Electrophoresis. Mar 1996;17(3):443-53. doi:10.1002/elps.1150170303
  • Referans2. Olivier M, Asmis R, Hawkins GA, Howard TD, Cox LA. The Need for Multi-Omics Biomarker Signatures in Precision Medicine. Int J Mol Sci. Sep 26 2019;20(19)doi:10.3390/ijms20194781
  • Referans3. Pandey A, Mann M. Proteomics to study genes and genomes. Nature. Jun 15 2000;405(6788):837-46. doi:10.1038/35015709
  • Referans4. Gramolini A, Lau E, Liu PP. Identifying Low-Abundance Biomarkers: Aptamer-Based Proteomics Potentially Enables More Sensitive Detection in Cardiovascular Diseases. Circulation. Jul 26 2016;134(4):286-9. doi:10.1161/CIRCULATIONAHA.116.022940
  • Referans5. Hanash S, Taguchi A. Application of proteomics to cancer early detection. Cancer J. Nov-Dec 2011;17(6):423-8. doi:10.1097/PPO.0b013e3182383cab
  • Referans6. Jiang S, Wang Y, Liu Z. The application of urinary proteomics for the detection of biomarkers of kidney diseases. Adv Exp Med Biol. 2015;845:151-65. doi:10.1007/978-94-017-9523-4_15
  • Referans7. Lin JL, Bonnichsen MH, Nogeh EU, Raftery MJ, Thomas PS. Proteomics in detection and monitoring of asthma and smoking-related lung diseases. Expert Rev Proteomics. Jun 2010;7(3):361-72. doi:10.1586/epr.10.9
  • Referans8. Aldred S, Grant MM, Griffiths HR. The use of proteomics for the assessment of clinical samples in research. Clinical Biochemistry. Nov 2004;37(11):943-952. doi:10.1016/j.clinbiochem.2004.09.002
  • Referans9. Conrotto P, Souchelnytskyi S. Proteomic approaches in biological and medical sciences: principles and applications. Exp Oncol. Sep 2008;30(3):171-80.
  • Referans10. Nakayasu ES, Gritsenko M, Piehowski PD, et al. Tutorial: best practices and considerations for mass-spectrometry-based protein biomarker discovery and validation. Nat Protoc. Aug 2021;16(8):3737-3760. doi:10.1038/s41596-021-00566-6
  • Referans11. Becker KF. Using tissue samples for proteomic studies-Critical considerations. Proteom Clin Appl. Apr 2015;9(3-4):257-267. doi:10.1002/prca.201400106
  • Referans12. Szklarczyk D, Franceschini A, Wyder S, et al. STRING v10: protein–protein interaction networks, integrated over the tree of life. Nucleic acids research. 2015;43(D1):D447-D452.
  • Referans13. Uslubas I, Kanli A, Kasap M, Akpinar G, Karabas L. Effect of aflibercept on proliferative vitreoretinopathy: Proteomic analysis in an experimental animal model. Exp Eye Res. Feb 2021;203doi:ARTN 108425 10.1016/j.exer.2020.108425
  • Referans14. Yoneten KK, Kasap M, Arga KY, Akpinar G, Utkan NZ. Decreased serum levels of glycerol-3- phosphate dehydrogenase 1 and monoacylglycerol lipase act as diagnostic biomarkers for breast cancer. Cancer Biomark. Oct 12 2021;doi:10.3233/CBM-203093
  • Referans15. Teke K, Kasap M, Simsek E, et al. SERPIN A5 may have a potential as a biomarker in reflecting the improvement of semen quality in infertile men who underwent varicocele repair. Andrologia. Aug 2021;53(7):e14081. doi:10.1111/and.14081
  • Referans16. Poli G, Ceni E, Armignacco R, et al. 2D-DIGE proteomic analysis identifies new potential therapeutic targets for adrenocortical carcinoma. Oncotarget. Mar 20 2015;6(8):5695-706. doi:10.18632/oncotarget.3299
  • Referans17. Lim MD, Dickherber A, Compton CC. Before You Analyze a Human Specimen, Think Quality, Variability, and Bias. Anal Chem. Jan 1 2011;83(1):8-13. doi:10.1021/ac1018974
  • Referans18. Espina V, Edmiston KH, Heiby M, et al. A portrait of tissue phosphoprotein stability in the clinical tissue procurement process. Mol Cell Proteomics. Oct 2008;7(10):1998-2018. doi:10.1074/mcp.M700596-MCP200
  • Referans19. Meric-Bernstam F, Akcakanat A, Chen H, et al. Influence of biospecimen variables on proteomic biomarkers in breast cancer. Clin Cancer Res. Jul 15 2014;20(14):3870-83. doi:10.1158/1078-0432.CCR-13-1507
  • Referans20. Gundisch S, Hauck S, Sarioglu H, et al. Variability of protein and phosphoprotein levels in clinical tissue specimens during the preanalytical phase. J Proteome Res. Dec 7 2012;11(12):5748-62. doi:10.1021/pr300560y
  • Referans21. Gundisch S, Grundner-Culemann K, Wolff C, et al. Delayed times to tissue fixation result in unpredictable global phosphoproteome changes. J Proteome Res. Oct 4 2013;12(10):4424-34. doi:10.1021/pr400451z
  • Referans22. Baker AF, Dragovich T, Ihle NT, Williams R, Fenoglio-Preiser C, Powis G. Stability of phosphoprotein as a biological marker of tumor signaling. Clin Cancer Res. Jun 15 2005;11(12):4338-40. doi:10.1158/1078-0432.CCR-05-0422
  • Referans23. Pinhel IF, Macneill FA, Hills MJ, et al. Extreme loss of immunoreactive p-Akt and p-Erk1/2 during routine fixation of primary breast cancer. Breast Cancer Res. 2010;12(5):R76. doi:10.1186/bcr2719
  • Referans24. Gao X, Yamazaki Y, Tezuka Y, et al. Intratumoral heterogeneity of the tumor cells based on in situ cortisol excess in cortisol-producing adenomas; similar to An association among morphometry, genotype and cellular senescence similar to. J Steroid Biochem. Nov 2020;204doi:ARTN 105764 10.1016/j.jsbmb.2020.105764
  • Referans25. Bertagna X, Guignat L, Groussin L, Bertherat J. Cushing's disease. Best Pract Res Clin Endocrinol Metab. Oct 2009;23(5):607-23. doi:10.1016/j.beem.2009.06.001
  • Referans26. Hatipoglu BA. Cushing's syndrome. J Surg Oncol. Oct 1 2012;106(5):565-71. doi:10.1002/jso.23197
  • Referans27. Newell-Price J, Bertagna X, Grossman AB, Nieman LK. Cushing's syndrome. Lancet. May 13 2006;367(9522):1605-17. doi:10.1016/S0140-6736(06)68699-6
  • Referans28. Nieman LK, Biller BM, Findling JW, et al. The diagnosis of Cushing's syndrome: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. May 2008;93(5):1526-40. doi:10.1210/jc.2008-0125
  • Referans29. Tabarin A, Perez P. Pros and cons of screening for occult Cushing syndrome. Nat Rev Endocrinol. Mar 22 2011;7(8):445-55. doi:10.1038/nrendo.2011.51
  • Referans30. Bansal V, El Asmar N, Selman WR, Arafah BM. Pitfalls in the diagnosis and management of Cushing's syndrome. Neurosurg Focus. Feb 2015;38(2):E4. doi:10.3171/2014.11.FOCUS14704
  • Referans31. Clayton RN, Raskauskiene D, Reulen RC, Jones PW. Mortality and morbidity in Cushing's disease over 50 years in Stoke-on-Trent, UK: audit and meta-analysis of literature. J Clin Endocrinol Metab. Mar 2011;96(3):632-42. doi:10.1210/jc.2010-1942
  • Referans32. Duan K, Hernandez KG, Mete O. Clinicopathological correlates of adrenal Cushing's syndrome. Postgrad Med J. Jun 2015;91(1076):331-42. doi:10.1136/postgradmedj-2014-202612rep
  • Referans33. Kjellin H, Johansson H, Hoog A, Lehtio J, Jakobsson PJ, Kjellman M. Differentially expressed proteins in malignant and benign adrenocortical tumors. PLoS One. 2014;9(2):e87951. doi:10.1371/journal.pone.0087951
  • Referans34. Yang MS, Wang HS, Wang BS, et al. A comparative proteomic study identified calreticulin and prohibitin up-regulated in adrenocortical carcinomas. Diagn Pathol. Apr 15 2013;8:58. doi:10.1186/1746-1596-8-58
  • Referans35. Kotlowska A, Malinski E, Sworczak K, Kumirska J, Stepnowski P. The urinary steroid profile in patients diagnosed with adrenal incidentaloma. Clin Biochem. Apr 2009;42(6):448-54. doi:10.1016/j.clinbiochem.2008.12.027
  • Referans36. Kotlowska A, Sworczak K, Stepnowski P. Urine metabolomics analysis for adrenal incidentaloma activity detection and biomarker discovery. J Chromatogr B Analyt Technol Biomed Life Sci. Feb 15 2011;879(5-6):359-63. doi:10.1016/j.jchromb.2010.12.021
  • Referans37. Arlt W, Biehl M, Taylor AE, et al. Urine steroid metabolomics as a biomarker tool for detecting malignancy in adrenal tumors. J Clin Endocrinol Metab. Dec 2011;96(12):3775-84. doi:10.1210/jc.2011-1565
  • Referans38. Kerkhofs TM, Kerstens MN, Kema IP, Willems TP, Haak HR. Diagnostic Value of Urinary Steroid Profiling in the Evaluation of Adrenal Tumors. Horm Cancer. Aug 2015;6(4):168-75. doi:10.1007/s12672-015-0224-3
  • Referans39. Minowada S, Kinoshita K, Hara M, Isurugi K, Uchikawa T, Niijima T. Measurement of urinary steroid profile in patients with adrenal tumor as a screening method for carcinoma. Endocrinol Jpn. Feb 1985;32(1):29-37. doi:10.1507/endocrj1954.32.29
  • Referans40. Stigliano A, Cerquetti L, Sampaoli C, Bucci B, Toscano V. Current and emerging therapeutic options in adrenocortical cancer treatment. J Oncol. 2012;2012:408131. doi:10.1155/2012/408131
  • Referans41. Tremoen NH, Fowler PA, Ropstad E, Verhaegen S, Krogenaes A. Exposure to the three structurally different PCB congeners (PCB 118, 153, and 126) results in decreased protein expression and altered steroidogenesis in the human adrenocortical carcinoma cell line H295R. J Toxicol Environ Health A. 2014;77(9-11):516-34. doi:10.1080/15287394.2014.886985
  • Referans42. Sbiera S, Leich E, Liebisch G, et al. Mitotane Inhibits Sterol-O-Acyl Transferase 1 Triggering Lipid-Mediated Endoplasmic Reticulum Stress and Apoptosis in Adrenocortical Carcinoma Cells. Endocrinology. Nov 2015;156(11):3895-908. doi:10.1210/en.2015-1367
  • Referans43. Kim HM, Lee YK, Koo JS. Proteome analysis of adrenal cortical tumors. Expert Rev Proteomics. Aug 2016;13(8):747-55. doi:10.1080/14789450.2016.1210008
  • Referans44. Sly WS, Hu PY. Human carbonic anhydrases and carbonic anhydrase deficiencies. Annu Rev Biochem. 1995;64:375-401. doi:10.1146/annurev.bi.64.070195.002111
  • Referans45. Supuran CT. Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat Rev Drug Discov. Feb 2008;7(2):168-81. doi:10.1038/nrd2467
  • Referans46. Ceylan Y, Akpinar G, Doger E, et al. Proteomic analysis in endometrial cancer and endometrial hyperplasia tissues by 2D-DIGE technique. J Gynecol Obstet Hum Reprod. Feb 2020;49(2):101652. doi:10.1016/j.jogoh.2019.101652
  • Referans47. Jo MS, Choi OH, Suh DS, et al. Correlation between expression of biological markers and [F]fluorodeoxyglucose uptake in endometrial cancer. Oncol Res Treat. 2014;37(1-2):30-4. doi:10.1159/000358163
  • Referans48. Sadlecki P, Bodnar M, Grabiec M, et al. The role of Hypoxia-inducible factor-1 alpha , glucose transporter-1, (GLUT-1) and carbon anhydrase IX in endometrial cancer patients. Biomed Res Int. 2014;2014:616850. doi:10.1155/2014/616850
  • Referans49. Petrova DT, Brehmer F, Schultze FC, et al. Differential kidney proteome profiling in a murine model of renal fibrosis under treatment with mycophenolate mofetil. Pathobiology. 2011;78(3):162-70. doi:10.1159/000324597
  • Referans50. Zahra K, Dey T, Ashish, Mishra SP, Pandey U. Pyruvate Kinase M2 and Cancer: The Role of PKM2 in Promoting Tumorigenesis. Front Oncol. 2020;10:159. doi:10.3389/fonc.2020.00159
  • Referans51. Duan K, Gucer H, Kefeli M, Asa SL, Winer DA, Mete O. Immunohistochemical Analysis of the Metabolic Phenotype of Adrenal Cortical Carcinoma. Endocr Pathol. Sep 2020;31(3):231-238. doi:10.1007/s12022-020-09624-3
  • Referans52. Warburg O, Wind F, Negelein E. The Metabolism of Tumors in the Body. J Gen Physiol. Mar 7 1927;8(6):519-30. doi:10.1085/jgp.8.6.519
Toplam 52 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Sağlık Kurumları Yönetimi
Bölüm Makaleler
Yazarlar

Aylin Kanlı 0000-0002-0674-0072

Turgay Şimşek 0000-0002-5733-6301

Murat Kasap Bu kişi benim 0000-0001-8527-2096

Gürler Akpınar 0000-0002-9675-3714

Zafer Cantürk 0000-0002-0042-9742

Proje Numarası KOÜ-BAP TSA-2020-1545
Yayımlanma Tarihi 30 Haziran 2022
Gönderilme Tarihi 2 Şubat 2022
Yayımlandığı Sayı Yıl 2022

Kaynak Göster

AMA Kanlı A, Şimşek T, Kasap M, Akpınar G, Cantürk Z. Tümör Doku Heterojenitesinin 2DE Proteomik Yaklaşımıyla Gösterilmesi. Sakarya Tıp Dergisi. Haziran 2022;12(2):301-314. doi:10.31832/smj.1066524

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