Gene Silencing in ovarian cancer cell line SKOV3 with siRNA using DreamFect Gold

Transient knockdown was performed using siRNA and DreamFect Gold transfection reagent.

This article demonstrates the high efficiency of DreamFect Gold from OZ Biosciences for gene silencing using siRNA in various cell lines.

article reference: Epigenomics, 2014, DOI:10.4161/15592294.2014.971608 

TGF-β induces global changes in DNA methylation during the epithelial-to-mesenchymal transition in ovarian cancer cells

Horacio Cardenas, Edyta Vieth, Jiyoon Lee, Mathew Segar, Yunlong Liu, Kenneth P. Nephew & Daniela Matei


Abstract

A key step in the process of metastasis is the epithelial-to-mesenchymal transition (EMT). We hypothesized that epigenetic mechanisms play a key role in EMT and to test this hypothesis we analyzed global and gene-specific changes in DNA methylation during TGF-β-induced EMT in ovarian cancer cells. Epigenetic profiling using the Infinium HumanMethylation450 BeadChip (HM450) revealed extensive (P < 0.01) methylation changes after TGF-β stimulation (468 and 390 CpG sites altered at 48 and 120 hour post cytokine treatment, respectively). The majority of gene-specific TGF-β-induced methylation changes occurred in CpG islands located in or near promoters (193 and 494 genes hypermethylated at 48 and 120 hours after TGF-β stimulation, respectively). Furthermore, methylation changes were sustained for the duration of TGF-β treatment and reversible after the cytokine removal. Pathway analysis of the hypermethylated loci identified functional networks strongly associated with EMT and cancer progression, including cellular movement, cell cycle, organ morphology, cellular development, and cell death and survival. Altered methylation and corresponding expression of specific genes during TGF-β-induced EMT included CDH1 (E-cadherin) and COL1A1 (collagen 1A1). Furthermore, TGF-β induced both expression and activity of DNA methyltransferases (DNMT) -1, -3A, and -3B, and treatment with the DNMT inhibitor SGI-110 prevented TGF-β-induced EMT. These results demonstrate that dynamic changes in the DNA methylome are implicated in TGF-β-induced EMT and metastasis. We suggest that targeting DNMTs may inhibit this process by reversing the EMT genes silenced by DNA methylation in cancer.

DreamFect Gold allows transfecting all types of nucleic acids with a very high efficiency. Due to its formulation, DreamFect™Gold delivers a large quantity of nucleic acids leading to higher protein expression compared to other transfection reagents. It is fully biodegradable and does not interfere with cellular mechanisms.