Inhibition of transcription factor T‐cell factor 3 (TCF3) using the oligodeoxynucleotide strategy increases embryonic stem cell stemness: possible application in regenerative medicine

Abstract

The transcription factor T‐cell factor 3 (TCF3), one component of the Wnt pathway, is known as a cell‐intrinsic inhibitor of many pluripotency genes in embryonic stem cells (ESCs) that influences the balance between pluripotency and differentiation. In this study, the effects of inhibition of TCF3 transcription factor on the stemness of mouse ESCs (mESCs) were investigated using the decoy oligodeoxynucleotides (ODNs) strategy. The TCF3 decoy and its scramble ODNs were designed and synthesized. The interaction specificity of the TCF3 decoy with the TCF3 transcription factor was evaluated by the electrophoretic mobility shift assay. Subcellular localization was carried out using fluorescence and confocal microscopy. Self‐renewal and pluripotency of mESCs were analyzed by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT), cell cycle and apoptosis, alkaline phosphatase (ALP), embryoid body (EB) formation, and real‐time assays. All experiments were performed in triplicate. The results showed that knockdown of TCF3 by decoy ODNs transfection in mESCs led to an increase in the cell proliferation, ALP enzyme activity, and master regulatory stemness genes and a decrease in the number and diameter of EBs. These results supported TCF3 as a potential target to maintain the pluripotency and self‐renewal capacity of mESCs. Knockdown of the TCF3 transcription factor using decoy ODNs can be a promising method to maintain the stemness of stem cells in regenerative medicine and cell therapy researches.

Suppressing the metastatic properties of the breast cancer cells using STAT3 decoy oligodeoxynucleotides: A promising approach for eradication of cancer cells by differentiation therapy

“…Transfection of Cy3‐labeled ODNs and Lipofectamine® 2000 resulted in high‐efficiency loading of ODNs (transfection rate >99%) into cells and significant mean fluorescent intensity by optimized increasing dose of ODNs, as shown in Figure a,b, from flow cytometry data, which further confirmed by fluorescent microscopy imaging. The results were consistent with our previous study (Johari et al, , ).…”

Evaluation of STAT3 decoy oligodeoxynucleotides' synergistic effects on radiation and/or chemotherapy in metastatic breast cancer cell line

“…Among several strategies, antisense therapy by oligodeoxynucleotides decoys (Bigdelou et al, 2019; Johari et al, 2019), which impairs interaction between transcription factors and related binding site in theirs correspond genes promoter in the genome, has been used successfully for blocking of the activities of STAT3 protein (Leong et al, 2003; Njatcha et al, 2018). This study aimed to evaluate the efficiency of a combinational therapy strategy of 22‐bp double‐stranded STAT3 decoy ODNs treatment with XI, MTX, and XI‐MTX exposure in highly metastatic breast cancer MDA‐MB‐231 cells through an in vitro analysis.…”

Application of decoy oligodeoxynucleotides strategy for inhibition of cell growth and reduction of metastatic properties in nonresistant and erlotinib‐resistant SW480 cell line

“…The novel therapeutic candidate, known as transcription factor decoys, could neutralize key transcription factors by mimicking their consensus DNA binding sites. Recent researches have shed light on the fact that decoy ODNs can competitively hinder the binding of the related TF to the target sequences on promoters (Johari et al, 2019; Rahmati et al, 2020). In this study, the designed STAT3 decoy ODNs were evaluated as a potential tool to suppress the STAT3 signaling pathway by blocking the phosphorylated STAT3 and downregulation of STAT3 downstream oncogenes.…”

“…Although, erlotinib inhibits effectively the EGFR, the progress of resistance to erlotinib during chemotherapy has led to treatment failure (Gharibi et al, 2020; Javadi et al, 2018; Weickhardt et al, 2012). These days, the combination of several therapeutic medications has become the main strategy for combating drug‐resistant cancers including RNAi (Chen et al, 2009; Roberts et al, 2017), antisense oligonucleotides (Preston et al, 2003), aptamers, ribozymes (Snir et al, 2003), and decoy oligodeoxynucleotides (Johari et al, 2019; Lasala & Minguell, 2011).…”