Lipopolisakkarit ve temozolomid ile muamele edilmiş glioblastoma hücresinde gremlin -1 ekspresyon düzeylerinin belirlenmesi
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Dosyalar
Tarih
2020
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Yayıncı
Tekirdağ Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Glioblastoma (GB) bilinen en maling beyin tümörüdür. Son yıllarda yapılan çalışmalar gremlin 1'i belirli kanser türlerinde potansiyel bir terapötik hedef olarak göstermektedir. Gremlin 1'in tümör dokularında kanserle ilişkili stromal hücreler tarafından eksprese edildiği ve bu şekilde mikroçevre tarafından regüle edilen tümör büyümesine ve invazyonuna katkıda bulunduğu düşünülmektedir. Glioblastoma Multiforma'nın kemoterapi tedavisinde sıklıkla temozolomid (TMZ) kullanılır ve anti-tümör etkisi olan tek alkilleyici ajandır. Lipopolisakkarit (LPS) tümör hücrelerini uyaran bir bakteri hücre duvar elemanı olup, gliomalar üzerinde anti-tümör etkisi bulunmaktadır. Bu deneysel çalışmada; U-118 insan glioblastoma multiforme hücre hattı, hücre kültürü ortamında çoğaltılarak, yaklaşık %95 yoğunluk gözlemlendiği gün flasktan hücreler tripsin-EDTA ile kaldırıldı. Hücre süspansiyonu hazırlanarak 60 mm2lik petrilere hücreler eklenerek çoğalması beklendi. İstenilen yoğunluğa ulaşan hücrelere 200 µM, 100µM, 50 µM, 25 µM konsantrasyon aralıklarında TMZ ve 10 µg/ml, 1 µg/ml, 0,1 µg/ml, 0,01 µg/ml konsantrasyon aralıklarında LPS uygulandı. 24 saat ve 48 saat inkübasyon sürelerinin hemen ardından protein izolasyonu yapıldı. Gremlin 1 protein düzeyleri sodyum dodesil sülfat-poliakrilamid jel elektroforezi (SDS-PAGE) ve Western blot ile analiz edildi. Çalışmamızda, 24 saat 25 ve 50 µM konsantrasyonlarda TMZ muamele edilen glioblastoma hücre hatlarında Gremlin-1 ekspresyonlarında artış ve 24 saat 0.1 µg/ml konsantrasyonda LPS uygulanan glioblastoma hücrelerinde gremlin-1 ifadesi seviyelerinde bir azalma gösterdik. Bu bulgular glioblastomada gremlin-1 düzeylerinin önemli bir rol oynadığına işaret etmektedir. Glioblastoma tümörlerinde direnç gelişme sıklığı ve sağ kalım süresinin kısalığı düşünüldüğünde gremlin-1'in etkinliğinin araştırılmasının oldukça önemli sonuçlar ortaya çıkarılabileceği kanısındayız.
Glioblastoma (GB) is the most known maling brain tumor. Recent studies have shown gremlin 1 as a potential therapeutic target for certain types of cancer. Gremlin 1 is thought to be expressed in tumor tissues by cancer-related stromal cells, thereby contributing to tumor growth and invasion regulated by microenvironment, and gremlin1 expression has been associated with tumor development. Temozolomide (TMZ) is often used in chemotherapy treatment of Glioblastoma Multiforme and is the only alkylating agent with an anti-tumor effect. Lipopolysaccharide is a bacterial cell wall element that stimulates tumor cells and has an anti-tumor effect on gliomas. In this experimental study; U-118 human glioblastoma multiforme cell line was replicated in cell culture medium, and cells were removed from the flask with trypsin-EDTA on the day when approximately 95% density was observed. The cell suspension was prepared and the cells were added to 60 mm2 petri dishes and they were allowed to multiply. The cells reaching the desired density were applied at 200 µM, 100 µM, 50 µM, 25 µM concentration ranges, TMZ and LPS at 10 µg / ml, 1 µg / ml, 0.1 µg / ml, 0.01 µg / ml concentration ranges. Protein isolation was performed immediately after the incubation times of 24 hours and 48 hours. Gremlin 1 protein levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. In our study, we showed an increase in gremlin-1 expressions in 24-hour 25 and 50 µM concentrations of TMZ-treated glioblastoma cell lines, and a decrease in Gremlin-1 expression in glioblastoma cells that received LPS at a concentration of 0.1 µg / ml for 24 hours. These findings indicate that gremlin-1 levels play an important role in glioblastoma. Considering the frequency of resistance development and shortness of survival in glioblastoma tumors, we believe that investigating the effectiveness of gremlin-1 can be quite significant.
Glioblastoma (GB) is the most known maling brain tumor. Recent studies have shown gremlin 1 as a potential therapeutic target for certain types of cancer. Gremlin 1 is thought to be expressed in tumor tissues by cancer-related stromal cells, thereby contributing to tumor growth and invasion regulated by microenvironment, and gremlin1 expression has been associated with tumor development. Temozolomide (TMZ) is often used in chemotherapy treatment of Glioblastoma Multiforme and is the only alkylating agent with an anti-tumor effect. Lipopolysaccharide is a bacterial cell wall element that stimulates tumor cells and has an anti-tumor effect on gliomas. In this experimental study; U-118 human glioblastoma multiforme cell line was replicated in cell culture medium, and cells were removed from the flask with trypsin-EDTA on the day when approximately 95% density was observed. The cell suspension was prepared and the cells were added to 60 mm2 petri dishes and they were allowed to multiply. The cells reaching the desired density were applied at 200 µM, 100 µM, 50 µM, 25 µM concentration ranges, TMZ and LPS at 10 µg / ml, 1 µg / ml, 0.1 µg / ml, 0.01 µg / ml concentration ranges. Protein isolation was performed immediately after the incubation times of 24 hours and 48 hours. Gremlin 1 protein levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. In our study, we showed an increase in gremlin-1 expressions in 24-hour 25 and 50 µM concentrations of TMZ-treated glioblastoma cell lines, and a decrease in Gremlin-1 expression in glioblastoma cells that received LPS at a concentration of 0.1 µg / ml for 24 hours. These findings indicate that gremlin-1 levels play an important role in glioblastoma. Considering the frequency of resistance development and shortness of survival in glioblastoma tumors, we believe that investigating the effectiveness of gremlin-1 can be quite significant.
Açıklama
Anahtar Kelimeler
Onkoloji, Oncology, Beyin neoplazmları, Brain neoplasms, Biyobelirteçler-tümör, Biomarkers-tumor, Gen ifadesi, Gene expression, Glioblastoma, Glioblastoma, Lipopolisakkaritler, Lipopolysaccharides, Neoplazmlar, Neoplasms, Temozolomid, Temosolomide
