title: The effects of shRNA-mediated gene silencing of transcription factor SNAI1 on the biological phenotypes of breast cancer cell line MCF-7.
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.
authors.affiliation: The Key Laboratory of Pathobiology, Ministry of Education, Department of Pathology, College of Basic Medical Sciences, Jilin University, 126 Xinmin Avenue, 130021, Changchun, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.forename: Lina
authors.initials: L
authors.lastname: Yu
authors.forename: Minlan
authors.initials: M
authors.lastname: Yang
authors.forename: Xiangshu
authors.initials: X
authors.lastname: Jin
authors.forename: Zhijing
authors.initials: Z
authors.lastname: Liu
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhang
authors.forename: Liping
authors.initials: L
authors.lastname: Wang
authors.forename: Dongjing
authors.initials: D
authors.lastname: Lin
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Chengshi
authors.initials: C
authors.lastname: Quan
mesh_terms: D016475:3T3 Cells; D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics; D001943:Breast Neoplasms / Q000145:classification / Q000235:genetics*; D015820:Cadherins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D062506:Claudin-4 / Q000096:biosynthesis; D057167:Claudins / Q000096:biosynthesis; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D061986:MCF-7 Cells; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis; D051379:Mice; D009361:Neoplasm Invasiveness / Q000235:genetics*; D010641:Phenotype; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D014408:Biomarkers, Tumor; C473255:CLDN7 protein, human; D015820:Cadherins; D062506:Claudin-4; D057167:Claudins; D034741:RNA, Small Interfering; C000604407:SNAI1 protein, human; C000604429:Snai1 protein, mouse; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2