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Role of Oct4–Sox2 complex decoy oligodeoxynucleotides strategy on reverse epithelial to mesenchymal transition (EMT) induction in HT29-ShE encompassing enriched cancer stem-like cells

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Abstract

Cancer stem cells are commonly tolerant toward chemotherapy and radiotherapy. Oct4 and Sox2 transcription factors are shown to be overexpressed in various cancers. At the current research, inhibition of Oct4 and Sox2 transcription factors was performed through application of decoy oligodeoxynucleotides (ODNs) strategy via repressing stemness properties in HT29-ShE cells encompassing enriched cancer stem-like cells. Designed Oct4–Sox2 complex decoy ODNs were transfected into HT29-ShE cells with Lipofectamine reagent. At the next step, ODNs efficiency transfection and subcellular localization were determined via flow cytometry and fluorescence microscopy, respectively. Further investigations such as cell proliferation and apoptosis analysis, colonosphere formation, invasion and migration, and real-time PCR assays were also carried out. Obtained results shed light on the fact that the designed complex decoys were effectively transfected into HT29-ShE cells, and they were found to be localized in subcellular compartments. Oct4–Sox2 decoy ODNs led to decreased cell viability, arresting the cell cycle in G0/G1 phases, increasing apoptosis, inhibition of migration/invasion and colonosphere formation ability of HT29-ShE cells in comparison with control and scramble groups. Furthermore, Oct4–Sox2 complex decoy could modulate the MET process via alteration of mRNA expression of downstream genes. It could be concluded that application of Oct4–Sox2 transcription factor decoy strategy in cells with stemness potential could lead to inhibiting the cell growth and triggering differentiation. Therefore, this technique could be applied along with usual remedies (chemotherapy and radiotherapy) as high potential method for treating cancer.

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Abbreviations

CSCs:

Cancer stem cells

cd‐ODNs:

Complex decoy oligodeoxynucleotides

EMSA:

Electrophoretic mobility shift assay

CRCs:

Colorectal cancer

MFI:

Mean fluorescence intensity

EMT:

Epithelial–mesenchymal transition

mESCs:

Mouse embryonic stem cells

HMG:

High‐mobility group

Oct4:

Octamer‐binding transcription Factor 4

Sox2:

Sex determining region Y‐box 2

TFD:

Transcription factor decoy

MET:

Mesenchymal–epithelial transition

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Acknowledgements

This study received support by Deputy for Research and Technology, Zanjan University of Medical Sciences, Zanjan, Iran (Grant Number A-12–1244-2, Ethical Code ZUMS.REC.1397.48).

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Authors and Affiliations

  1. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Zahra Bigdelou, Yousef Mortazavi, Zoleykha Asadi & Behrooz Johari

  2. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

    Yousef Mortazavi & Behrooz Johari

  3. Department of Genetics, Breast Cancer Research Center, Motamed Cancer Institute, Academic Center for Education, Culture and Research, Tehran, Iran

    Zohreh Saltanatpour

  4. Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran

    Mehdi Kadivar

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  1. Zahra Bigdelou
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  2. Yousef Mortazavi
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  3. Zohreh Saltanatpour
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  4. Zoleykha Asadi
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Contributions

Conception of Idea and study design: ZB, and BJ. Collecting data, and paper writing: ZB, BJ, MK, ZS, and ZA. Final approval: YM and BJ.

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Correspondence to Behrooz Johari.

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Bigdelou, Z., Mortazavi, Y., Saltanatpour, Z. et al. Role of Oct4–Sox2 complex decoy oligodeoxynucleotides strategy on reverse epithelial to mesenchymal transition (EMT) induction in HT29-ShE encompassing enriched cancer stem-like cells. Mol Biol Rep 47, 1859–1869 (2020). https://doi.org/10.1007/s11033-020-05280-2

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