Onur TOKGÜN, Nedim KARAGENÇ, Pervin Elvan TOKGÜN, Kubilay INCI, Hakan AKÇA, Gamze GÖKÖZ DOĞU, Aydın DEMİRAY

KRAS mutasyonlarının kolon ve akciğer kanseri eksozomlar aracılığıyla tayini ve primer dokuya kıyasla mutasyon durumlarının analizi

Amaç: Kolorektal kanser ve akciğer kanserleri olan hastalarda epidermal büyüme faktörü reseptör hedefli tedavinin etkinliği önemli ölçüde KRAS mutasyonu ile ilişkilidir. Bu çalışmanın amacı, Kolorektal kanser ve akciğer kanserli hastaların serum ekzozom ve primer tümör dokusunda ki KRAS mutasyon durumlarının karşılaştırmaktır. Gereç ve Yöntemler: Histolojik olarak doğrulanmış 19 adet kolorektal kanser ve 28 adet akciğer kanserli toplam 47 hastanın tümör dokularından genomik DNA izole edildi ve iki kür tedavi sonrasında ilgili hastalardan alınan periferik kandan ekzomal RNA izole edildi. KRAS geninde üç bölgede gözlenen mutasyonlar (kodon 12, 13 ve 61) pyrosekanslama yöntemi ile analiz edildi. Elde edilen sekans verileri doğrultusunda tümör dokuları ile serum ekozomlarında bulunan nükleik asitlere ait KRAS mutasyon durumları ortaya kondu. Bulgular: Doku örneklerinden KRAS mutasyon profilleri belirlenen hastaların eksozomlarında gözlenen KRAS mutasyonları incelendiğinde 3 (%6,38) hastada dokuda belirlenen mutasyonun dışında yeni bir mutasyon saptanmış, 9 (%19,14) hastada doku örneğinde mutasyon var iken herhangi bir KRAS mutasyonu saptanamamış, 5 (%10,6) hastada doku örneğinde mutasyon yok iken herhangi bir KRAS mutasyonu saptanmıştır. 30 (%63,8) hastada ise mutasyon olup olmama durumu doku ve eksozom analizleri doğrultusunda değişmemiştir. Sonuç: Elde ettiğimiz sonuçlar, kolorektal kanser ve akciğer kanserli hastaların hızlı ve non-invaziv bir materyal ile genotiplenmesinde serum ekzomal mRNA'nın yeni ve güvenilir bir kaynak olarak kullanılabileceği ileri sürmektedir.

Anahtar Kelimeler:

Likit biyopsi, KRAS, eksozom, DNA

Detection of KRAS mutations by colon and lung cancer exosomes and analysis of mutational statuses compared to primary tissue

Aim: The efficacy of epidermal growth factor receptor-targeted therapy in colorectal cancer and lung cancers is significantly associated with the KRAS mutation. Therefore, this study aims to compare the KRAS mutation status in serum exosome and primary tumor tissue of colorectal and lung cancer patients. Material and Methods: Genomic DNA was isolated from tumor tissues of 47 patients with histologically confirmed 19 colorectal cancers and 28 lung cancers, and exosomal RNA was isolated from peripheral blood from the respective patients after two treatment cycles. Mutations (codons 12, 13, and 61) observed in three regions of the KRAS gene were analyzed by pyrosequencing. The sequence data obtained revealed the KRAS mutation status of nucleic acids in tumor tissues and serum exosomes. Results: When the KRAS mutations were observed in the exosomes of the cases whose KRAS mutation profiles were determined from the tissue samples, a new mutation other than the mutation determined in the tissue was detected in 3 (6.38%) cases. In comparison, there was a mutation in the tissue sample in 9 (19.14%) cases, and no KRAS mutation could be detected in 5 (10.6%) cases. While there was no mutation in the tissue sample, any KRAS mutation was detected. In 30 (63.8%) cases, the presence or absence of mutation did not change in line with tissue and exosome analysis. Conclusion: Our results suggest that serum exosomal mRNA can be used as a new and reliable source for genotyping patients with colorectal and lung cancer with a rapid and non-invasive material.

Keywords:

liquid biopsy, KRAS, exosome, DNA,

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