Expression of miR-92, miR-21, miR-155, Oncogemic miRNAs using qRT-PCR in Colon Cancer stage II Tissues

Öz

Background: In this study, we aimed to detect the expression levels of miR-92, miR-21 and miR-155 in tumor and normal colon and rectum tissues with oncogenic microRNA (miRNA) obtained from patients with Stage II colorectal cancer using qRTPCR and to compare the obtained data. Material and Method: This study was carried out with the clinical, laboratory and pathological findings of the tissues obtained from patients diagnosed with colorectal cancer at Gaziantep University Medical Faculty Research and Training Hospital -General Surgery Department. miRNeasy mini kit-isolated miRNAs were used for qRT-PCR implementation. The isolated miRNAs were converted into cDNA via the miScript RT-PCR commercial kit. The cDNA samples with miRNA expression levels in tissue samples were identified via Rotor Gene RT-PCR system using RT-SYBR Green qPCR kit. Result: The mean level of miRNA-21 expression in 9 patients diagnosed with colorectal cancer appears to have increased 2.09 fold compared to normal tissue and tumor tissue, while miR-155 increased 2.22 fold. This increase was considered as statistically significant (p <0.05). When the fold chance values of normal and tumor tissue are compared, despite being oncogenic miRNA, miR-92 appears to have decreased 1.37 fold and was not considered statistically significant (p>0.05). Conclusion: According to the findings obtained in our study, the expression levels of oncogenic miRNA miR21 and miR-155 in tumor colon tissues of patients with colorectal cancer have increased. The results of recent studies have shown that miRNAs involved in the regulation of many cellular functions may be used as biomarkers for diagnosis and treatment of cancer.

Anahtar Kelimeler

• miRNA
• oncogenic miRNA
• colorectal cancer
• biomarker

Stage II Kolon kanseri dokularında Onkogenik microRNA olan miR-92, miR21, miR-155’in qRT-PCR’da Ekspresyonu

Öz

Amaç: Bu çalışmada Stage II kolorektal kanserli hastalardan alınmış tümörlü ve normal kolon/rektum dokularında Onkogenik mikroRNA (miRNA) olan miR-92, miR-21 ve miR-155’in ekspresyon seviyelerinin qRT-PCR ile tespit edilmesi ve elde edilen verilerin karşılaştırılması amaçlanmıştır. Materyal ve Metod: Bu çalışma Gaziantep Üniversitesi Tıp Fakültesi Araştırma ve Uygulama Hastanesi Genel Cerrahi Anabilim Dalı’ndan laboratuar ve patalojik bulgular ile klinik değerlendirme sonucunda kolorektal kanser tanısı konmuş 9 hastanın tümörlü ve normal (kontrol-temiz cerrahi sınırları) kolon/rektum dokular ile çalışılmıştır. qRT-PCR uygulaması için miRNeasy mini kit ile izole edilen miRNA’lar kullanıldı. İzole edilen miRNA’lar miScript RT-PCR ticari kit ile cDNA’ya dönüştürüldü. Doku örneklerindeki miRNA ekspresyon seviyeleri ile cDNA örnekleri RT- SYBR Green qPCR kiti kullanılarak Rotor Gene RT-PCR sistemi ile belirlendi. Bulgular: Kolorektal kanser teşhisi konulmuş 9 hastanın ortalama miRNA-21 ekspresyon seviyesi normal doku ve tümörlü doku karşılaştırıldığında 2.09 kat arttığı, miR-155’in ise 2.22 kat arttığı tespit edilmiştir. Bu artış istatiksel olarak anlamlı bulunmuştur (p≤ 0.05). onkogenik miRNA olmasına rağmen miR-92 ekpresyon miktarı normal ve tümör dokusu fold change değerleri karşılaştırıldığında 1.37 kat azaldığı tespit edilmiştir. Ancak bu azalmanın istatiksel bakımdan anlamlı olmadığı bulunmuştur (p˃0.05). Sonuç: Çalışmamızdan elde ettiğimiz bulgulara göre kolorektal kanserde onkogenik miRNA olan miR-21 ve miR-155’in hastaların tümörlü kolon dokularında ekspresyon seviyeleri artmıştır. Son yıllarda yapılan araştırmaların sonuçları Hücresel birçok temel işlevin düzenlenmesinde görev alan miRNA’ların kanserin tanı ve tedavisi için biyobelirteç olarak kullanılabileceği fikrini ortaya koymuştur.

Anahtar Kelimeler

• miRNA
• onkogenik miRNA
• Kolon kanseri
• Biyobelirteç

Kaynakça

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