Primer ve Metastatik Melanom Hücre Hatlarında Sıcak Nokta Mutasyonlarının Karşılaştırılması

Öz

Amaç: Melanom, UV radyasyonunun mutajenik bir risk faktörü olarak hareket etmesi sonucu gelişen ve çeşitli genetik değişikliklere neden olan bir hastalıktır. Çalışmamızda Primer Melanom (WM115) ve Metastatik Melanom (SK-MEL-30) hücre hatları arasında insan kanser ilişkili genlerindeki değişiklikleri belirlemeyi amaçladık. Materyal ve Metot: Proto-onkogenler ve tümör baskılayıcı genler arasından insan kanserleriyle ilişkili 50 gen seçildi. Daha sonra iki hücre hattı arasında 50 gende oluşan hotspot mutasyonlarının değişimleri Ion Ampliseq Cancer Hotspot DNA Panel v2 gen paneli kullanılarak incelendi. Bulgular: Metastatik melanom hücre hattında primer hücre hattına göre mutasyon artışı gözlendi. WM115 hücre hattında 4 adet Önemi Belirsiz Varyant (VUS), 3 adet benign ve 2 adet patojenik mutasyon tespit edildi. BRAF proto-onkogeninde patojenik mutasyonlar tanımlanmıştır. SK-MEL-30 hücre hattında 24 VUS, 1 iyi huylu ve 1 olası patojenik mutasyon (VHL geninde) gözlendi. Sonuç: Araştırmamızın sonuçlarına göre, primer melanomda BRAF mutasyonunun görülmesinin, bu genin melanomun ilerlemesi ve gelişiminde etkili olduğunu, metastatik melanomda ise VHL geninde oluşan mutasyonun ise HIF yolu ile karsinogenezde etkili olabileceğini düşünmekteyiz. Ek olarak, metastatik melanomun VUS mutasyonlarının çoğu RET ve NOTCH yollarında meydana geldi. Bu yollarda yer alan genlerdeki mutasyonların birikmesi, metastaz gelişimine katkıda bulunabilir.

Anahtar Kelimeler

• Hotspot mutasyonu
• melanom
• NOTCH
• RET
• yeni nesil dizileme

Destekleyen Kurum

Mevcut çalışma, Ankara Yıldırım Beyazit Üniversitesi Proje Koordinasyon Uygulama ve Araştırma Merkezi (hibe numarası 18) tarafından desteklenmiştir.

Proje Numarası

Ankara Yildirim Beyazit University Project Coordination Application and Research Center (grant no. 18).

Etik Beyan

Çalışma için etik onay gerekmemektedir. Bu çalışmada ikincil hücre kültürü kullanıldı.

The Comparison of Hotspot Mutations in Primary and Metastatic Melanoma Cell Lines

Öz

Objective: Melanoma is a disease that develops as a result of UV radiation acting as a mutagenic risk factor and causes various genetic changes. In our study, we aimed to determine the changes in human cancer-related genes between Primary Melanoma (WM115) and Metastatic Melanoma (SK-MEL-30) cell lines. Materials and Methods: Among the proto-oncogenes and tumor suppressor genes, 50 genes associated with human cancers were selected. Afterwards, the changes of hotspot mutations occurring in 50 genes between the two cell lines were examined using the Ion Ampliseq Cancer Hotspot DNA Panel v2 gene panel. Results: A mutation increase was observed in the metastatic melanoma cell line compared to the primary cell line. 4 Variant of Uncertain Significance (VUS), 3 benign and 2 pathogenic mutations were detected in the WM115 cell line. Pathogenic mutations have been identified in the BRAF proto-oncogene. In the SK-MEL-30 cell line, 24 VUS, 1 benign, and 1 possible pathogenic mutation (in the VHL gene) were observed. Conclusions: According to the results of our research, we think that the occurrence of BRAF mutations in primary melanoma indicates that this gene is effective in the progression and development of melanoma, and the mutation occurring in the VHL gene of metastatic melanoma might be effective in carcinogenesis via the HIF pathway. In addition, most of the VUS mutations of metastatic melanoma occurred in the RET and NOTCH pathways. The accumulation of mutations in genes involved in these pathways may contribute to the development of metastasis.

Anahtar Kelimeler

• Hotspot mutation
• melanoma
• next generation sequencing
• NOTCH
• RET

Destekleyen Kurum

The present study was supported by the Ankara Yildirim Beyazit University Project Coordination Application and Research Center (grant no. 18).

Proje Numarası

Ankara Yildirim Beyazit University Project Coordination Application and Research Center (grant no. 18).

Etik Beyan

Ethical approval for the study is not required. In this study, secondary cell culture was used.

Kaynakça

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