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The effects of shRNA-mediated gene silencing of transcription factor SNAI1 on the biological phenotypes of breast cancer cell line MCF-7

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Abstract

To research the effects of silencing transcription factor SNAI1 on the in vitro biological phenotypes of breast cancer cell line MCF-7, based on the gene sequence of SNAI1, we linked shRNA with the green fluorescent protein-expressing eukaryotic expression vector pGCsilencer™ U6/Neo/GFP, and transfected it into MCF-7 cells. The SNAI1 gene-silencing effect was authenticated by RT-PCR and immunofluorescence. We then examined the effect of gene silencing on the expression of epithelial and mesenchymal markers and on their biological phenotypes of the target cells. Finally, we explained that SNAI1 was bound to E-cadherin in MCF-7 cells by ChIP. Silencing SNAI1 upregulated the expression of epithelial markers claudin-4, claudin-7, and E-cadherin, while expression of the mesenchymal marker matrix metalloproteinase-2 was downregulated. The capacity for proliferation, migration, and invasion was diminished. SNAI1 binds to the E-cadherin gene promoter and inhibits its transcription. We can conclude that silencing gene SNAI1 inhibits expression of properties that are associated with the malignant phenotype of MCF-7 cells and reverses the epithelial–mesenchymal transition process by regulating relevant target gene E-cadherin.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (Code: 81172499) and Science and Technology Development Plan of the Office of Science and Technology Project in Jilin Province (Code: 201115113). We thank Dr. William Orr, Department of Pathology, University of Manitoba, Canada, for his help in the preparation of the manuscript.

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Authors and Affiliations

  1. The Key Laboratory of Pathobiology, Ministry of Education, Department of Pathology, College of Basic Medical Sciences, Jilin University, 126 Xinmin Avenue, 130021, Changchun, China

    Yan Lu, Minlan Yang, Xiangshu Jin, Zhijing Liu, Xiaowei Zhang, Liping Wang, Dongjing Lin, Yuanyuan Liu, Min Wang & Chengshi Quan

  2. The First Hospital Cancer Center of Jilin University, Changchun, China

    Lina Yu

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  1. Yan Lu
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  2. Lina Yu
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  3. Minlan Yang
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Correspondence to Chengshi Quan.

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Yan Lu and Lina Yu have contributed equally to this paper.

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Lu, Y., Yu, L., Yang, M. et al. The effects of shRNA-mediated gene silencing of transcription factor SNAI1 on the biological phenotypes of breast cancer cell line MCF-7. Mol Cell Biochem 388, 113–121 (2014). https://doi.org/10.1007/s11010-013-1903-4

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