Akciğer kanserli hastalarda periferik kandan ctDNA'nın metilasyon değişikliklerinin dijital PCR ile incelenmesi
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Dosyalar
Tarih
2020
Yazarlar
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Yayıncı
Ege Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Akciğer kanseri, dünya genelinde en sık görülen ve her iki cinsiyette de en sık ölüme sebep olan malignitelerden biridir. En sık sebebi sigara dumanı olan akciğer kanserinin erken tanısında, sigara içenlerde düşük doz bilgisayarlı tomografi ile tarama önerilmiştir. Bu taramanın yanlış tanı, maliyet, zaman kaybı gibi dezavantajları nedeniyle akciğer kanserinin erken tanısında non invaziv, sensitif biyomarkerların kullanılmasına ihtiyaç duyulmuştur. Bu çalışmada, ileri evre-küçük hücre dışı (non-small cell) akciğer kanseri tanısı konulmuş hastaların plazmasından elde edilen cfDNA'da (cell-free DNA) literatür taraması ile belirlenmiş akciğer kanseri ile ilişkisi bilinen 6 gene ait (RASSF1A, p16, SHOX2, RARβ2, APC, DAPK1) droplet dijital PCR (ddPCR) ölçümü ile saptanan metilasyon değişikliklerinin sağlıklı kişilerdeki değerleri ile karşılaştırarak değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem: Çalışmaya alınan 10 hasta ve 10 kontrolün periferik kan plazma örneklerinden cfDNA elde edildi. Literatür taraması ile 6 hedef gen promotor bölgesi belirlendi. cfDNA'da hedef gen bölgelerinin bisülfit dönüşümü sonrasında ddPCR ile metilasyon analizi yapıldı. Çıkan sonuçlar istatistiksel olarak analiz edildi. Bulgular: Çalışmada DAPK1 geninin promotor bölgesindeki metilasyon seviyelerinde, hasta ve kontrol grubu arasında anlamlı farklılık saptanmıştır (p<0,05). Çalışmaya alınan 10 hastanın 8'inde DAPK1 geninde anlamlı metilasyon yüksekliği saptanmıştır. DAPK1 genindeki hipermetilasyon literatür ile uyumlu bulunmuştur. Sonuç: Çalışmamız ddPCR ile akciğer kanserinde cfDNA'dan metilasyon analizi yapan ülkemizdeki öncü çalışmalardan biri olmuştur. Küçük hücre dışı akciğer kanserinde DAPK1 geninde hipermetilasyon saptanması ile literatüre katkıda bulunmuştur. Çalışmamız, bu alanda daha fazla sayıda hastalarla yapılacak sonraki çalışmalar için ön çalışma niteliğindedir.
Objective: Lung cancer is one of the most common malignancies worldwide and the most common cause of death in both sexes. The most common cause of lung cancer is cigarette smoke. In the early diagnosis of lung cancer in smokers, screening with low-dose computed tomography has been proposed. Due to the disadvantages of this screening such as misdiagnosis, cost and time loss; non-invasive and sensitive biomarkers were needed for early diagnosis of lung cancer. In this study, it was aimed to evaluate the methylation changes detected by droplet digital PCR (ddPCR) measurement of 6 genes (RASSF1A, p16, SHOX2, RARβ2, APC, DAPK1) known to be associated with lung cancer determined by the literature scan in cfDNA (cell-free DNA) obtained from the plasma of patients diagnosed with advanced-non-small cell lung cancer. Materials and Methods: cfDNA was obtained from peripheral blood plasma samples of 10 patients and 10 controls included in the study. With the literature review, 6 target gene promoter regions were determined. After bisulfite transformation of target gene regions in cfDNA, methylation analysis was performed with ddPCR. The results were analyzed statistically. Results: In the study, a significant difference was found between the patient and control groups in the methylation levels of the DAPK1 gene in the promoter region (p <0,05). In 8 of 10 patients included in the study, significant methylation height in the DAPK1 gene was detected. Hypermethylation in the DAPK1 gene has been found to be compatible with the literature. Conclusion: Our study has been one of the leading studies in our country that perform methylation analysis from cfDNA in lung cancer with ddPCR. It contributed to the literature with the detection of hypermethylation in the DAPK1 gene in non-small cell lung cancer. Our study is a preliminary study for further studies in this field with more patients.
Objective: Lung cancer is one of the most common malignancies worldwide and the most common cause of death in both sexes. The most common cause of lung cancer is cigarette smoke. In the early diagnosis of lung cancer in smokers, screening with low-dose computed tomography has been proposed. Due to the disadvantages of this screening such as misdiagnosis, cost and time loss; non-invasive and sensitive biomarkers were needed for early diagnosis of lung cancer. In this study, it was aimed to evaluate the methylation changes detected by droplet digital PCR (ddPCR) measurement of 6 genes (RASSF1A, p16, SHOX2, RARβ2, APC, DAPK1) known to be associated with lung cancer determined by the literature scan in cfDNA (cell-free DNA) obtained from the plasma of patients diagnosed with advanced-non-small cell lung cancer. Materials and Methods: cfDNA was obtained from peripheral blood plasma samples of 10 patients and 10 controls included in the study. With the literature review, 6 target gene promoter regions were determined. After bisulfite transformation of target gene regions in cfDNA, methylation analysis was performed with ddPCR. The results were analyzed statistically. Results: In the study, a significant difference was found between the patient and control groups in the methylation levels of the DAPK1 gene in the promoter region (p <0,05). In 8 of 10 patients included in the study, significant methylation height in the DAPK1 gene was detected. Hypermethylation in the DAPK1 gene has been found to be compatible with the literature. Conclusion: Our study has been one of the leading studies in our country that perform methylation analysis from cfDNA in lung cancer with ddPCR. It contributed to the literature with the detection of hypermethylation in the DAPK1 gene in non-small cell lung cancer. Our study is a preliminary study for further studies in this field with more patients.
Açıklama
Anahtar Kelimeler
KüçüK Hücre Dışı Akciğer Kanseri, cfDNA, DAPK1, ddPCR, Non-small Cell Lung Cancer
