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| IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711 |
Int J Clin Exp Med 2013;6(4):259-268
Original Article
Expression levels of microRNA machinery components Drosha, Dicer and DGCR8 in
human (AGS, HepG2, and KEYSE-30) cancer cell lines
Naser Jafari, Hadi Peeri Dogaheh, Shahab Bohlooli, Glenn G Oyong, Zohreh Shirzad, Fatemeh Alibeiki, Saeid Hosseini Asl, Seyed
Jalal Zargar
Department of Biochemistry, School of Medicine, Ardabil University of Medical Science, Ardabil 56197, Iran; Department of Cell &
Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran; Department of Microbiology, School of
Medicine, Ardabil University of Medical Science, Ardabil 56197, Iran; Department of Pharmacology, School of Medicine, Ardabil
University of Medical Science, Ardabil 56197, Iran; Molecular Science Unit, Center for Natural Science and Ecological Research, and
Biology Department, De La Salle Uni-versity, Manila, Philippines
Received March 15, 2013; Accepted March 30, 2013; Epub April 12, 2013; Published April 30, 2013
Abstract: MicroRNAs (miRNAs) have recently been shown to play fundamental roles in diverse cellular processes and linked to variety
of cancers. Dicer and Drosha are two major enzymes in the miRNA maturation process. DGCR8 is the assistant of Drosha in the
microprocessor complex. In this study, we evaluated the mRNA expression profiles of major miRNA processing machinery Drosha,
Dicer, and DGCR8 in human gastrointestinal (AGS, KYSE30 and HepG2) cancer cell lines. Materials and Methods: The cells were
cultured and harvested, and total cellular RNA was isolated from cells. Then, first-strand cDNA was synthesized from the RNA of cells.
Afterward, Quantitative analysis was performed by real-time RT-PCR using the PowerSYBR Green PCR Master Mix. Results:
Expression levels of Drosha in AGS and HepG2 cells were higher than the controls, whereas, Drosha’s expression level in KYSE-30
cell line was lower. The Dicer expression levels in AGS and HepG2 cells were higher, while, its expression level in KYSE-30 cell was
lower. The DGCR8 expression levels in all three cell lines were significantly higher than the control samples. Conclusion: Expression
levels of the two most important enzymes of the miRNA machinery, Drosha and Dicer, and microprocessor complex component,
DGCR8 were noticeably dysregulated when compared to healthy controls. (IJCEM1303010).
Keywords: Drosha, Dicer, DGCR8, microRNA machinery, cancer cell line
Address correspondence to: Dr. Naser Jafari, Department of Biochemistry, School of Medicine, Ardabil University of Medical Sciences,
Ardabil 56197, Iran. Phone: +98 (451) 5522089; Email: n.jafari@arums.ac.ir
Original Article
Expression levels of microRNA machinery components Drosha, Dicer and DGCR8 in
human (AGS, HepG2, and KEYSE-30) cancer cell lines
Naser Jafari, Hadi Peeri Dogaheh, Shahab Bohlooli, Glenn G Oyong, Zohreh Shirzad, Fatemeh Alibeiki, Saeid Hosseini Asl, Seyed
Jalal Zargar
Department of Biochemistry, School of Medicine, Ardabil University of Medical Science, Ardabil 56197, Iran; Department of Cell &
Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran; Department of Microbiology, School of
Medicine, Ardabil University of Medical Science, Ardabil 56197, Iran; Department of Pharmacology, School of Medicine, Ardabil
University of Medical Science, Ardabil 56197, Iran; Molecular Science Unit, Center for Natural Science and Ecological Research, and
Biology Department, De La Salle Uni-versity, Manila, Philippines
Received March 15, 2013; Accepted March 30, 2013; Epub April 12, 2013; Published April 30, 2013
Abstract: MicroRNAs (miRNAs) have recently been shown to play fundamental roles in diverse cellular processes and linked to variety
of cancers. Dicer and Drosha are two major enzymes in the miRNA maturation process. DGCR8 is the assistant of Drosha in the
microprocessor complex. In this study, we evaluated the mRNA expression profiles of major miRNA processing machinery Drosha,
Dicer, and DGCR8 in human gastrointestinal (AGS, KYSE30 and HepG2) cancer cell lines. Materials and Methods: The cells were
cultured and harvested, and total cellular RNA was isolated from cells. Then, first-strand cDNA was synthesized from the RNA of cells.
Afterward, Quantitative analysis was performed by real-time RT-PCR using the PowerSYBR Green PCR Master Mix. Results:
Expression levels of Drosha in AGS and HepG2 cells were higher than the controls, whereas, Drosha’s expression level in KYSE-30
cell line was lower. The Dicer expression levels in AGS and HepG2 cells were higher, while, its expression level in KYSE-30 cell was
lower. The DGCR8 expression levels in all three cell lines were significantly higher than the control samples. Conclusion: Expression
levels of the two most important enzymes of the miRNA machinery, Drosha and Dicer, and microprocessor complex component,
DGCR8 were noticeably dysregulated when compared to healthy controls. (IJCEM1303010).
Keywords: Drosha, Dicer, DGCR8, microRNA machinery, cancer cell line
Address correspondence to: Dr. Naser Jafari, Department of Biochemistry, School of Medicine, Ardabil University of Medical Sciences,
Ardabil 56197, Iran. Phone: +98 (451) 5522089; Email: n.jafari@arums.ac.ir
