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The transcriptional factor Snail simultaneously triggers cell cycle arrest and migration of human hepatoma HepG2

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

Snail was recently highlighted as a critical transcriptional factor for tumor metastasis. Real time RT/PCR and Western blot analysis demonstrated that Snail mRNA and protein, respectively, were induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in hepatoma cell HepG2. Blockade of gene expression of Snail by antisense oligodeoxynucleotide and/or siRNA technique can prevent not only the TPA-triggered EMT/cell migration and growth inhibition of HepG2 but also TPA-induced down-regulation of E-cadherin and up-regulation of p15INK4b. Moreover, the TPA-triggered promoter activation of p15INK4b was also prevented. On the other hand, two of the HepG2 clone over-expressing Snail, namely S7 and S15, had a scattered fibroblastic morphology and acquired higher motility than parental HepG2. Also, the proportion of G0/G1 phase of S7 and S15 was higher than that of parental HepG2, consistent with the longer doubling time of both cells. Semiquantitative RT/PCR analysis demonstrated a greatly elevated gene expression of Snail accompanied with decreased E-cadherin and increased p15INK4b in both Snail-overexpressing cells. On the transcriptional level, p15INK4b promoter activity was 2.6-fold higher in S7 as compared with parental HepG2. Furthermore, electrophoretic mobility of DNA fragments encompassing proximal p15INK4b promoter can be retarded by incubation of nuclear extract of S7. Our results demonstrated that Snail play diverse trans-regulatory roles in HepG2. Notably, we suggested that Snail may upregulate p15INK4b gene expression by directly activating its promoter.

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Abbreviations

EMT:

Epithelial mesenchymal transition

TPA:

Tumor promoter 12-O-tetradecanoylphorbol-13-acetate

EMSA:

Electrophoretic mobility shift assay

S/AS ODNs:

Sense/antisense oligodeoxynucleotides

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Acknowledgement

We thank National Science Council in Taiwan for financial support, Dr. Ji-Hshiung Chen in Tzu Chi University for providing the Snail expressing plasmid and Mr. Shih S. M. for technical assistance.

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Authors and Affiliations

  1. Research Centre for Hepatology, Department of Internal Medicine, Buddhist Tzu Chi General Hospital and Tzu Chi University, Hualien, Taiwan

    Chi-Tan Hu

  2. Department of Medical Technology, Tzu Chi University, No. 701, Chung Yang Rd, Sec 3, Hualien, 970, Taiwan

    Jia-Ru Wu, Tsu Yao Chang, Chuan-Chu Cheng & Wen-Sheng Wu

  3. Graduate Institute of Medicine, Tzu Chi University, Hualien, Taiwan

    Jia-Ru Wu, Tsu Yao Chang, Chuan-Chu Cheng & Wen-Sheng Wu

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  1. Chi-Tan Hu
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  2. Jia-Ru Wu
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  5. Wen-Sheng Wu
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Correspondence to Wen-Sheng Wu.

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Hu, CT., Wu, JR., Chang, T.Y. et al. The transcriptional factor Snail simultaneously triggers cell cycle arrest and migration of human hepatoma HepG2. J Biomed Sci 15, 343–355 (2008). https://doi.org/10.1007/s11373-007-9230-y

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  • DOI: https://doi.org/10.1007/s11373-007-9230-y

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