Araştırma Makalesi
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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

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.

Kaynakça

  • 1) Wijnhoven B.P, Michael M.Z and Watson D.I. MicroRNAs and cancer. BR. Journel Surgery 2007;94:23-30. 2) Iorio M.V, Ferracin M, Liu C.G, et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005; 65:7065–7070. 3) Calin G.A, Ferracin M, Cimmino A, Di Leva G, Shimizu M, Wojcik SE, et al. A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. The New England journal of medicine 2005; 353(17):1793-801. 4) Lehmann U, Hasemeier B, Römermann D, Müler K, Langer F. and Kreipe H., Epigenetic inactivation of microRNA genes in mammary carcinoma, Verh Dtsch Ges Pathol 200;7 91, 214- 220 p. 5) Saydam F, Değirmenci İ, Güneş V. MikroRNA’lar ve kanser Dicle Tıp Dergisi, 2011; 38(1), 113-120. 6) Michor F, Iwasa Y, et al. Dynamics of colorectal cancer. Semin Cancer Biol. 2005; 15(6), 484-93. 7) Hermsen M, Postma C and Baak J. Colorectal adenoma to carcinoma progression follows multiple pathways of chromosomal instability. Gastroenterolgy 2002; 123, 1109-1119. 8) Corte H, Manceau G, Blons H, Laurent-Puig P. MicroRNA and colorectal cancer. Dig Liver Dis. 2012; 195-200. 9) Sigel R.L, Miller K.B, Jemal A. Cncer Statistic, 2017. Cancer Journal for Clinicions 2017; 67: 7- 30 10) Ambros V. A hierarchy of regulatory genes controls a larva-to-adult developmental switch in C. elegans. Cell 1989; 57(1):49-57. 11) Lee R.C, Feinbaum R.L, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 1993; 75(5):843 54. 12) Aiello M, Vella N, Cannavo C, Scalisi A, Spandidos D.A, Toffoli C, Buonadonna A, Libra M, and Stivala F. Role of genetic polymorphisms and mutations in colorectal cancer therapy. Molecular Medicine Reports 2011; 4, 203-208. 13) Bedeir A and Krasinskas A.M. Molecular Diagnostics of Colorectal Cancer. Arch Pathol Lab Med 2011; 135, 204-215. 14) Pillai R.S, Bhattacharyya S.N, Filipowicz W. Repression of protein synthesis by miRNAs: How many mechanisms? Trends in Cell Biology 2007; 17, 118-126. 15) Calin G. A et al. Frequent deletions and downregülation of microRNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proceedings of the National Academy of Sciences of the United States of America 2002; 99:15524- 15529. 16) Calin G.A, Sevignani C, Dumitru C.D, et al. Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA 2004;101:2999– 3004. 17) Hayes J, Peruzzi P.P and Lawler S. microRNAs in cancer: biomarkers, functions and therapy. Trends in Molecular Medicine 2014; 20(8). 18) Mazeh H, Mizrahi I, Ilyayev N, Halle D, Brücher B, et al. The Diagnostic and Prognostic Role of microRNA in Colorectal Cancer-a Comprehensive review. J Cancer 2013; 4: 281-295. 19) Shen J, Stass S.A, Jiang F. MicroRNAs as potential biomarkers in human solid tumors. Cancer Lett 2013; 329: 125-136. 20) Michael M.Z, Connor S.M, and et al. “Reduced accumulation of specific microRNAs in colorectal neoplasia”, Mol Cancer Res 2003; 1, 882-891 21) Baraniskin A, Birkenkap K, and Maghnoui A. “MiR-30a- 5p suppresses tumor growth in colon carcinoma by targeting DTL”, Carcinogenesis 2012; 33, 732-739. 22) Schetter A.J, Okayama H, Harris C. The Role of MicroRNAs in Colorectal Cancer. Cancer J 2012; 18, 244-252. 23) Callari M, Dugo M, Musella V, Marchesi E, Chiorini G, et al. Comparison of Microarray Platforms for Measuring Differential MicroRNA Expression in Paired Normal/Cancer Colon Tissues. PLoS ONE 2012; 7(9), e45105. 24) Ye J.J, Cao J. MicroRNAs in colorectal cancer as markers and targets: Recent advances. World J Gastroenterol 2014 20(15), 4288-4299 25) Manne U, Shanmugam C, Bovell L, et al.: miRNAs as biomarkers for management ofpatients with colorectal cancer. Biomark Med 2010; 4:761–770. 26) Slaby O, Svoboda M, Fabian P, et al. Altered expression of miR-21, miR-31, miR-143 and miR145 is related to clinicopathologic features of colorectal cancer. Oncology. 2008; 72(5-6):397– 402. 27) Huang J, Zheng S, Jin SH, Zhang SZ. Somatic mutations of APC gene in carcinomas from hereditary non-polyposis colorectal cancer patients. World J Gastroenterol 2004; 10: 834-836. 28) Wang J.C, Zhou Z.G, Wang L, et al. Clinicopathological significance of microRNA31, -143 and -145 expression in colorectal cancer. Disease Markers. 2009; 26(1):27–34

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

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

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.

Kaynakça

  • 1) Wijnhoven B.P, Michael M.Z and Watson D.I. MicroRNAs and cancer. BR. Journel Surgery 2007;94:23-30. 2) Iorio M.V, Ferracin M, Liu C.G, et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005; 65:7065–7070. 3) Calin G.A, Ferracin M, Cimmino A, Di Leva G, Shimizu M, Wojcik SE, et al. A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. The New England journal of medicine 2005; 353(17):1793-801. 4) Lehmann U, Hasemeier B, Römermann D, Müler K, Langer F. and Kreipe H., Epigenetic inactivation of microRNA genes in mammary carcinoma, Verh Dtsch Ges Pathol 200;7 91, 214- 220 p. 5) Saydam F, Değirmenci İ, Güneş V. MikroRNA’lar ve kanser Dicle Tıp Dergisi, 2011; 38(1), 113-120. 6) Michor F, Iwasa Y, et al. Dynamics of colorectal cancer. Semin Cancer Biol. 2005; 15(6), 484-93. 7) Hermsen M, Postma C and Baak J. Colorectal adenoma to carcinoma progression follows multiple pathways of chromosomal instability. Gastroenterolgy 2002; 123, 1109-1119. 8) Corte H, Manceau G, Blons H, Laurent-Puig P. MicroRNA and colorectal cancer. Dig Liver Dis. 2012; 195-200. 9) Sigel R.L, Miller K.B, Jemal A. Cncer Statistic, 2017. Cancer Journal for Clinicions 2017; 67: 7- 30 10) Ambros V. A hierarchy of regulatory genes controls a larva-to-adult developmental switch in C. elegans. Cell 1989; 57(1):49-57. 11) Lee R.C, Feinbaum R.L, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 1993; 75(5):843 54. 12) Aiello M, Vella N, Cannavo C, Scalisi A, Spandidos D.A, Toffoli C, Buonadonna A, Libra M, and Stivala F. Role of genetic polymorphisms and mutations in colorectal cancer therapy. Molecular Medicine Reports 2011; 4, 203-208. 13) Bedeir A and Krasinskas A.M. Molecular Diagnostics of Colorectal Cancer. Arch Pathol Lab Med 2011; 135, 204-215. 14) Pillai R.S, Bhattacharyya S.N, Filipowicz W. Repression of protein synthesis by miRNAs: How many mechanisms? Trends in Cell Biology 2007; 17, 118-126. 15) Calin G. A et al. Frequent deletions and downregülation of microRNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proceedings of the National Academy of Sciences of the United States of America 2002; 99:15524- 15529. 16) Calin G.A, Sevignani C, Dumitru C.D, et al. Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA 2004;101:2999– 3004. 17) Hayes J, Peruzzi P.P and Lawler S. microRNAs in cancer: biomarkers, functions and therapy. Trends in Molecular Medicine 2014; 20(8). 18) Mazeh H, Mizrahi I, Ilyayev N, Halle D, Brücher B, et al. The Diagnostic and Prognostic Role of microRNA in Colorectal Cancer-a Comprehensive review. J Cancer 2013; 4: 281-295. 19) Shen J, Stass S.A, Jiang F. MicroRNAs as potential biomarkers in human solid tumors. Cancer Lett 2013; 329: 125-136. 20) Michael M.Z, Connor S.M, and et al. “Reduced accumulation of specific microRNAs in colorectal neoplasia”, Mol Cancer Res 2003; 1, 882-891 21) Baraniskin A, Birkenkap K, and Maghnoui A. “MiR-30a- 5p suppresses tumor growth in colon carcinoma by targeting DTL”, Carcinogenesis 2012; 33, 732-739. 22) Schetter A.J, Okayama H, Harris C. The Role of MicroRNAs in Colorectal Cancer. Cancer J 2012; 18, 244-252. 23) Callari M, Dugo M, Musella V, Marchesi E, Chiorini G, et al. Comparison of Microarray Platforms for Measuring Differential MicroRNA Expression in Paired Normal/Cancer Colon Tissues. PLoS ONE 2012; 7(9), e45105. 24) Ye J.J, Cao J. MicroRNAs in colorectal cancer as markers and targets: Recent advances. World J Gastroenterol 2014 20(15), 4288-4299 25) Manne U, Shanmugam C, Bovell L, et al.: miRNAs as biomarkers for management ofpatients with colorectal cancer. Biomark Med 2010; 4:761–770. 26) Slaby O, Svoboda M, Fabian P, et al. Altered expression of miR-21, miR-31, miR-143 and miR145 is related to clinicopathologic features of colorectal cancer. Oncology. 2008; 72(5-6):397– 402. 27) Huang J, Zheng S, Jin SH, Zhang SZ. Somatic mutations of APC gene in carcinomas from hereditary non-polyposis colorectal cancer patients. World J Gastroenterol 2004; 10: 834-836. 28) Wang J.C, Zhou Z.G, Wang L, et al. Clinicopathological significance of microRNA31, -143 and -145 expression in colorectal cancer. Disease Markers. 2009; 26(1):27–34
Toplam 1 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Klinik Tıp Bilimleri
Bölüm Araştırma Makalesi
Yazarlar

Çiğdem Güngörmez

Hatice Gumushan Aktas

Ersin Borazan Bu kişi benim

Yayımlanma Tarihi 30 Aralık 2017
Gönderilme Tarihi 30 Ekim 2017
Kabul Tarihi 22 Kasım 2017
Yayımlandığı Sayı Yıl 2017 Sayı: 3 - IONCC 2017 Özel sayısı

Kaynak Göster

Vancouver Güngörmez Ç, Aktas HG, Borazan E. Stage II Kolon kanseri dokularında Onkogenik microRNA olan miR-92, miR21, miR-155’in qRT-PCR’da Ekspresyonu. Harran Üniversitesi Tıp Fakültesi Dergisi. 2017;14(3):31-9.

Harran Üniversitesi Tıp Fakültesi Dergisi  / Journal of Harran University Medical Faculty