Publication: Tümör mikroçevresinde ve tümörde etkin yolakların glioma seyrindeki rolü
Abstract
Amaç: Glial hücrelerin farklılaşması sonucu oluşan gliomada bir çok yolak etkilidir. Hücre proliferasyonunda etkili olan Hippo yolağıdır bunlardan biridir. Çalışmamızın amacı HGG ve LGG dokularında ve mikro çevresinde Hippo yolağı üzerinden metastazda etkin LATS2 (Large Tumor Suppressor Kinase 2 ve proliferasyonda etkili olan WWTR1 (WW Domain Containing Transcription Regulator) ve hücre adezyonunda etkili aşağı yöndeki CYR61 (Cysteine-rich angiogenic inducer 61) genlerinin ekspresyonlarını karşılaştırmalı olarak saptamaktır. Gereç ve Yöntem: Çalışmaya Marmara Üniversitesi Eğitim Araştırma Hastanesi Beyin ve Sinir Cerrahisi AD’ de opere olan beş adet LGG (yaş: 47,2±17,9) ve sekiz adet HGG (yaş: 60±10,06) hastası dahil edildi. Dokulardan mRNA izolasyonu ardından cDNA’ya sentezi gerçekleştirildi. RT-qPCR yöntemiyle SYBR green kullanılarak yapılan çalışmada hedef genlerin ekspresyonları BioRad CFX 96 cihazında analiz edildi. İç kontrol olarak ACTB (B Actin) geni kullanıldı. Genlerin HGG’lerde LGG’lere oranla kat artışları 2^–∆∆Ct metoduyla GeneGlobe Software’de analiz edildi. Bulgular: LGG ve HGG örneklerinin gen ekspresyonları birbirlerine göre analiz edildiğinde CYR61’in ekspresyon değerleri HGG örneklerinde 3,09 kat yüksek bulundu. LATS2 ve WWTR1’in ekspresyonlarında anlamlılık saptanmadı. GBM tümörü ve tümör mikroçevresinin ekspresyonları karşılaştırıldığında tümörde LATS2’nin ekspresyonu azalırken CYR61 ve WWTR1’in arttığı saptandı. Yapılan regresyon analizlerinde ve demografik olgularda da istatiksel anlamlı sonuçlar elde edilmedi. Sonuç: CYR61, epitel-mezenkimal dönüşümü sağlayarak olarak glioma gelişimine neden olur. Çalışmamızda HGG hastalarımızda saptanan bu bulgu, tümörün GBM’e dönüşebilme kapasitesi olduğunu göstermektedir. Tümörde olduğu gibi tümör mikroçevresinde de gen ekspresyon analizlerinin yapılması klinik seyir hakkında bilgi verebilecek önemli bir analiz aracı olmalıdır.
Objective: Many pathways are effective in glioma formed because of differentiation of glial cells. Proliferation related Hippo pathway is one of them. Objective of our study is to express genes of LATS2(Large Tumor Suppressor Kinase 2), which effects metastasis, WWTR1(WW Domain Containing Transcription Regulator) that changes proliferation, and downstream CYR61(Cysteine-rich angiogenic inducer 61), which alters cell adhesion in HGG&LGG tissues and microenvironment via Hippo pathway. Material and Methods: Five LGG (age: 47.2±17.9) and eight HGG (age: 60±10.06) patients who were operated in Marmara University Training and Research Hospital, Department of Neurosurgery were included. mRNA isolation and cDNA synthesis were performed. SYBR green with RT-qPCR method was used, expressions of genes were analyzed in BioRad CFX 96 device. ACTB (B Actin) gene was used as internal control. Fold increase of genes in HGGs compared to LGGs was analyzed with 2^–∆∆Ct method. Results: When gene expressions of LGG&HGG samples were analyzed, expression values of CYR61 were found 3.09 times higher in HGGs. Expressions of LATS2 and WWTR1 were not significant. When expressions of GBM tumor and tumor microenvironment were compared, it was found that expression of LATS2 in tumor decreased, while CYR61 and WWTR1 increased. No statistically significant results were obtained. Conclusion: CYR61 leads glioma by providing epithelial-mesenchymal transformation. This finding, which was found in our study, shows that tumor has capacity to transform into GBM. Gene expression analyzes in tumor microenvironment as well as in the tumor should be an important analysis tool that can provide information about clinical course.
Objective: Many pathways are effective in glioma formed because of differentiation of glial cells. Proliferation related Hippo pathway is one of them. Objective of our study is to express genes of LATS2(Large Tumor Suppressor Kinase 2), which effects metastasis, WWTR1(WW Domain Containing Transcription Regulator) that changes proliferation, and downstream CYR61(Cysteine-rich angiogenic inducer 61), which alters cell adhesion in HGG&LGG tissues and microenvironment via Hippo pathway. Material and Methods: Five LGG (age: 47.2±17.9) and eight HGG (age: 60±10.06) patients who were operated in Marmara University Training and Research Hospital, Department of Neurosurgery were included. mRNA isolation and cDNA synthesis were performed. SYBR green with RT-qPCR method was used, expressions of genes were analyzed in BioRad CFX 96 device. ACTB (B Actin) gene was used as internal control. Fold increase of genes in HGGs compared to LGGs was analyzed with 2^–∆∆Ct method. Results: When gene expressions of LGG&HGG samples were analyzed, expression values of CYR61 were found 3.09 times higher in HGGs. Expressions of LATS2 and WWTR1 were not significant. When expressions of GBM tumor and tumor microenvironment were compared, it was found that expression of LATS2 in tumor decreased, while CYR61 and WWTR1 increased. No statistically significant results were obtained. Conclusion: CYR61 leads glioma by providing epithelial-mesenchymal transformation. This finding, which was found in our study, shows that tumor has capacity to transform into GBM. Gene expression analyzes in tumor microenvironment as well as in the tumor should be an important analysis tool that can provide information about clinical course.