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Negative Regulatory Role of TWIST1 on SNAIL Gene Expression

  • Original Article
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Pathology & Oncology Research

Abstract

Epithelial-mesenchymal transition (EMT) is crucial for specific morphogenetic movements during embryonic development as well as pathological processes of tumor cell invasion and metastasis. TWIST and SNAIL play vital roles in both developmental and pathological EMT. Our aim in this study was to investigate the functional correlation between TWIST1 and SNAIL in human ESCC cell line (KYSE-30). The packaging cell line GP293T was cotransfected with either control retroviral pruf-IRES-GFP plasmid or pruf-IRES-GFP-hTWIST1 and pGP plasmid. The KYSE-30 ESCC cells were transduced with produced viral particles and examined with inverted fluorescence microscope. DNA was extracted from transduced KYSE-30 cells and analyzed for copy number of integrated retroviral sequences in the target cell genome. The concentration of retroviral particles was determined by Real-time PCR. After RNA extraction and cDNA synthesis, the mRNA expression of TWIST1 and SNAIL was assessed by comparative real-time PCR amplification. Ectopic expression of TWIST1 in KYSE-30, dramatically reduces SNAIL expression. Retroviral transduction enforced TWIST1 overexpression in GFP-hTWIST1 nearly 9 folds in comparison with GFP control cells, and interestingly, this TWIST1 enforced expression caused a − 7 fold decrease of SNAIL mRNA expression in GFP-hTWIST1 compared to GFP control cells. Inverse correlation of TWIST1 and SNAIL mRNA levels may introduce novel molecular gene expression pathway controlling EMT process during ESCC aggressiveness and tumorigenesis. Consequently, these data extend the spectrum of biological activities of TWIST1 and propose that therapeutic repression of TWIST1 may be an effective strategy to inhibit cancer cell invasion and metastasis.

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Acknowledgments

The authors gratefully acknowledge the colleagues at the Division of Human Genetics for their technical assistance. This study was supported by a grant (No.91046360) from Iran National Science Foundation (INSF).

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Authors and Affiliations

  1. Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran

    Mohammad Mahdi Forghanifard & Abolfazl Rad

  2. Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Bu-Ali square, Mashhad, 9196773117, Iran

    Sima Ardalan Khales & Mohammad Reza Abbaszadegan

  3. Molecular Medicine Research Department, ACECR-Khorasan Razavi Branch, Mashhad, Iran

    Moein Farshchian

  4. Department of Biochemistry and Biophysics, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Masoud Homayouni-Tabrizi

Authors
  1. Mohammad Mahdi Forghanifard
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  2. Sima Ardalan Khales
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  3. Moein Farshchian
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  4. Abolfazl Rad
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  5. Masoud Homayouni-Tabrizi
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  6. Mohammad Reza Abbaszadegan
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Correspondence to Mohammad Reza Abbaszadegan.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Mohammad Mahdi Forghanifard and Sima Ardalan Khales are contributed equally in this study.

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Forghanifard, M.M., Ardalan Khales, S., Farshchian, M. et al. Negative Regulatory Role of TWIST1 on SNAIL Gene Expression. Pathol. Oncol. Res. 23, 85–90 (2017). https://doi.org/10.1007/s12253-016-0093-2

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  • DOI: https://doi.org/10.1007/s12253-016-0093-2

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