Expression of miR-143, miR-145, miR-192, Tumor Suppressor miRNAs using qPCR in Colon Cancer stage II

Öz

Background: In this study, it was aimed to determine the expression levels of tumor suppressor microRNAs (miRNA) including miR-143, miR-145 and miR-192 in tumor and normal colon / rectum tissues of Stage II colorectal cancer (CRC) patients by using qPCR and to compare the obtained data. Materials and Methods: This study was performed on tumor or normal (control-clean surgical margin) colon / rectum tissues of 9 patients diagnosed with colorectal cancer as a result of clinical evaluation with laboratory and pathological findings in Gaziantep University Medical Faculty Research and Practice Hospital General Surgery Department. miRNAs isolated with miRNeasy mini kit were used for qPCR. Isolated miRNAs were converted into cDNA by miScript RTPCR commercial kit. The miRNA expression levels of tissue samples were determined by using RT- SYBR Green qPCR kit in Rotor Gene Q. Result: The mean level of miRNA-143 expression in 9 patients diagnosed with colorectal cancer appears to have decreased 0.096 fold compared to normal tissue and tumor tissue, while miR-145, 1.52 fold. This decrease was considered as statistically significant (p <0.05). When the fold chance values of normal and tumor tissue are compared, despite being tumor supressor miRNA, miR-192 appears to have increased 3.25 fold and was not considered statistically significant (p>0.05). Conclusion: According to the findings obtained in our study, the expression levels of tumor supressor miRNA miR-143 and miR-145 in tumor colon tissues of patients with colorectal cancer have decreased. miRNAs have attracted major interest as a means to analyze the molecular pathways involved in cancer development and progression. In addition to their important cellular functions, it is possible that secreted miRNAs may be diagnostic biomarkers for cancer detection. 

Anahtar Kelimeler

• miRNA
• Tumor suppressor miRNA
• Biomarker
• Colorectal cancer

Stage II Kolon Kanserinde Tümör Süpressör miRNA olan miR-143, miR-145 ve miR-192’nin Ekspresyonu

Öz

Amaç: Bu çalışmada Stage II kolorektal kanserli hastalardan alınmış tümörlü ve normal kolon/rektum dokularında Tümör süpressör mikroRNA (miRNA) olan miR-143, miR-145 ve miR-192’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’nda laboratuar ve patalojik bulgular ile klinik değerlendirme sonucunda kolorektal kanser tanısı konmuş 9 hastanın tümörlü ve normal (kontrol-temiz cerrahi sınırları) kolon/rektum dokuları ile gerçekleştirilmiştir. qRTPCR 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 RT- SYBR Green qPCR kiti kullanılarak Rotor Gene Q sistemi ile belirlendi. Bulgular: Kolorektal kanser teşhisi konulmuş 9 hastanın ortalama miRNA-143 ekspresyon seviyesi normal doku ve tümörlü doku karşılaştırıldığında 0.096 kat azaldığı, miR-145’in ise 1.52 kat azaldığı tespit edilmiştir. Bu artış istatiksel olarak anlamlı bulunmuştur (p˂0,05). miR-192 tümör baskılayıcı miRNA olmasına rağmen ekpresyon miktarının normal ve tümör dokusu fold chance değerleri karşılaştırıldığında 3.25 kat arttığı tespit edilip istatiksel olarak anlamlı bulunmamıştır (p˃0,05). Sonuç: Çalışmamızdan elde ettiğimiz bulgulara göre kolorektal kanserde tümör süpressör miRNA olan miR-143 ve miR-145’in hastaların tümörlü kolon dokularında ekspresyon seviyeleri azalmıştır. miRNA'lar, kanser gelişimi ve ilerlemesindeki moleküler yollarda önemli etkiler etmektedir. miRNA’lar hücresel birçok işleve ek olarak, kanser tanı ve tedavisinde biyobelirteç olarak kullanılabileceği düşünülmektedir.

Anahtar Kelimeler

• miRNA
• Tümör baskılayıcı miRNA
• Biyobelirteç
• Kolon kanseri

Kaynakça

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