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Volume 7, Issue 3(Suppl) J Biotechnol Biomater, an open access journal
ISSN: 2155-952X Euro Biotechnology 2017
September 25-27, 2017
17th EURO BIOTECHNOLOGY CONGRESS
September 25-27, 2017 Berlin, Germany
Study of the role of siRNAmediated promoter methylation in DNMT3B knockdown and alteration of promoter
methylation of CDH1, GSTP1 genes in MDA-MB -453 cell line
Mojgan Naghitorabi1, Hamid Mir Mohammad Sadeghi2, Javad Mohammadi Asl 1, Mohammad Rabbani2 and Abbas Jafarian-Dehkordi 2
1Ahvaz Jundishapur University of Medical Sciences, Iran
2Isfahan University of Medical Sciences, Iran
Promoter methylation is one of the main epigenetic mechanisms that leads to the inactivation of tumor suppressor genes during
carcinogenesis. Due to the reversible nature of DNA methylation, many studies have been performed to correct theses epigenetic
defects by inhibiting DNA methyltransferases (DNMTs). In this case novel therapeutics especially siRNA oligonucleotides have been
used to specifically knock down the DNMTs at mRNA level. Also many studies have focused on transcriptional gene silencing in
mammalian cells via siRNA mediated promoter methylation. The present study was designed to assess the role of siRNA mediated
promoter methylation in DNMT3B knockdown and alteration of promoter methylation of Cadherin-1 (CDH1), Glutathione
S-Transferase Pi 1(GSTP1), and DNMT3B genes in MDA-MB-453 cell line. MDA-MB-453 cells were transfected with siDNMT
targeting DNMT3B promoter and harvested at 24 and 48 h post transfection to monitor gene silencing and promoter methylation.
DNMT3B expression was monitored by quantitative RT-PCR method. Promoter methylation was quantitatively evaluated using
differential high resolution melting analysis. A non-significant 20% reduction in DNMT3B mRNA level was shown only after first
transfection with siDNMT. Promoter methylation levels of DNMT3B, CDH1, and GSTP1 were detected at about 15%, 70% and 10%
respectively, in the MDA-MB-453 cell line, with no significant change after transfection.Our results indicated that siDNMT sequence
were not able to affect promoter methylation and silencing of DNMT3B in MDA-MB-453 cells. However, quantitation of methylation
confirmed a hypermethylated phenotype at CDH1 and GSTP1 promoters as well as a differential methylation pattern at DNMT3B
promoter in breast cancer.
Biography
Mojgan Naghitorabi is an Assistant Professor of the School of Pharmacy at Ahvaz Jundishapur University of Medical Sciences, Iran. She received her Pharm D and
PhD degrees from the School of Pharmacy and Pharmaceutical Sciences at Isfahan University of Medical Sciences, Iran. Her research interests lie in the area of
epigenetics, regulation of gene expression, RNA interference, and cancer. She has published four papers in her research field.
Mojgan Naghitorabi et al., J Biotechnol Biomater 2017, 7:3(Suppl)
DOI: 10.4172/2155-952X-C1-077