The Demonstration of Tumor Heterogeneity using 2DE Proteomic Approach

Abstract

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.

Keywords

• Tumor heterogeneity
• Proteomics
• Cushing Syndrome

Tümör Doku Heterojenitesinin 2DE Proteomik Yaklaşımıyla Gösterilmesi

Abstract

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.

Keywords

• Tümör heterojenitesi
• Proteomik
• Cushing Sendromu

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