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

Yıl 2017, Sayı: 3 - IONCC 2017 Özel sayısı, 31 - 39, 30.12.2017

Çiğdem Güngörmez , Hatice Gumushan Aktas , Ersin Borazan

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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