The importance of FISH Test Targeting EGFR, CCND1 and RREB1 Genes in Differentiating Malignant Melanomas from Melanocytic Nevus

Öz

Melanocytic nevus (MN) may on occasion be difficult to distinguish from malignant melanoma (MM) histopathologically. Fluorescent in situ hybridization (FISH) has been demonstrated to be of use for the diagnosis of melanocytic neoplasms of the skin. In this study, the effectiveness of the standart melanoma FISH test (4-way probe targeting RREB1, CCND1, MYB genes and centromere 6) and additionally probes, targetting EGFR, TP53, MDM2 and TP53 genes, in differentiating melanomas from melanocytic nevi were investigated. Standard FISH test was performed on 24 MM and 24 MN samples, but EGFR, TP53, MDM2 and TP53 gene copy numbers were investigated in 16 of 24 MM and 24 MN using FISH method. The incidence of FISH-detected positive genomic copy aberrations (4-way probe, and others) was determined as 83,3% in 24 MM cases, and 5,2% in 24 MN. Statistically significant differences were found between the MM and MN groups in terms of CCND1, RREB1, EGFR amplifications (p<0.001, p<0.05, p˂0.05), but there was no association between histopathological features and detected abnormalities (p>0,05). In additionally, all 5 acral lentiginous melanomas, could be analysed, had EGFR amplifications. In conclusion, CCND1, RREB1, and EGFR amplifications have diagnostic significance for MM. The FISH test is very effective in terms of its use as an adjunct to histopathological methods. But centromere controlled probes should be used to avoid false positive results.

Anahtar Kelimeler

• FISH
• EGFR
• CCND1
• RREB1
• Malignant melanoma

Malign Melanomlar ile Melanositik Nevüs Ayrımında EGFR, CCND1 ve RREB1 Genlerini Hedef Alan FISH Testinin Yeri

Öz

Melanositik nevüs (MN) örneklerini histopatolojik olarak malign melanomdan (MM) ayırmak bazen zor olabillmektedir. Floresan in situ hibridizasyon (FISH)’in ciltteki melanositik neoplazmların teşhisinde kullanıldığı kanıtlanmıştır. Bu çalışmada standart melanom FISH testinin (RREB1, CCND1, MYB genlerini ve sentromer 6'yı hedefleyen 4 yollu prob) ve ayrıca EGFR, TP53, MDM2 ve TP53 genlerini hedefleyen probların melanomları melanositik nevüslerden ayırmadaki etkinliği araştırılmıştır. 24 MM ve 24 MN örneklerine standart FISH testi uygulandı. Ancak EGFR, TP53, MDM2 ve TP53 gen kopya sayıları FISH yöntemi kullanılarak 24 MN ve 16/24 MM örneklerinde incelendi. Floresan in situ hibridizasyon ile saptanan pozitif genomik kopya aberasyonlarının (4 yollu prob ve diğerleri) görülme sıklığı 24 MM vakasında %83,3 ve 24 MN'de %5,2 olarak belirlendi. Malign melanom ve MN grupları arasında CCND1, RREB1, EGFR amplifikasyonları açısından istatistiksel olarak anlamlı farklılıklar bulundu (p<0,001, p<0,05, p˂0,05), ancak histopatolojik özellikler ile saptanan anormallikler arasında ilişki saptanmadı (p>0,05). Ek olarak, 5 akral lentijinöz melanomun tümü analiz edilebildi ve hepsi EGFR amplifikasyonu açısından pozitifti. Sonuç olarak, CCND1, RREB1 ve EGFR amplifikasyonları MM için tanısal öneme sahip olduğu ve FISH testinin, histopatolojik yöntemlere ek olarak kullanılmasının etkili olabileceği sonucuna varılmıştır. Ancak, yanlış pozitif sonuçlardan kaçınmak için sentromer kontrollü problar kullanılması gerekliliği gözlenmiştir.

Anahtar Kelimeler

• FISH
• EGFR
• CCND1
• RREB1
• malign melanom

Kaynakça

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