title: Snail-induced epithelial-mesenchymal transition in gastric carcinoma cells and generation of cancer stem cell characteristics.
abstract: Biological changes in Snail-overexpressed SGC7901 cells were studied by establishing a pEGFP-C1-Snail carrier. The significance of Snail in epithelial-mesenchymal transition (EMT) as well as the invasion and metastatic capacity of gastric cancer cells was also discussed; moreover, we attempted to verify the probable cancer stem cell characteristics of Snail-overexpressed cells. A pEGFP-C1-Snail eukaryotic expression plasmid was constructed and pEGFP-C1(-) and pEGFP-C1-Snail plasmids were extracted and transfected into SGC7901 cells using Lipofectamine 2000. Stably expressed SGC7901-N [control group containing pEGFP-C1(-)] and SGC7901-S (test group containing pEGFP-C1-Snail) cells were screened using a G418 resistance medium. Snail, E-cadherin, b-catenin, vimentin, and fibronectin gene and protein expressions were detected by real-time quantitative PCR, western blot, and immunofluorescence. Cell invasion and metastasis were tested by scratch test, invasion assay, and an adhesion experiment. The positive rate of aldehyde dehydrogenase-1 (ALDH-1) expression was analyzed by flow cytometry. The results indicated the occurrence of EMT, accompanied by morphological changes in the cells and a weakening of the cell adhesion capacity. We also observed a decrease in the expression of epithelial markers E-cadherin and b-catenin and an increase in mesenchymal (Snail and vimentin) marker expression. Moreover, the cells showed increased invasiveness and metastatic capacity, and decreased proliferative ability. Moreover, the Snail-treated SGC7901 cells moved towards the scratch and produced fewer clones compared to the control cells. Owing to its capacity for self-renewal, SGC7901-S cells produced new clones and expressed ALDH-1. Therefore, we concluded that Snail overexpression induced EMT and endowed cells with tumor stem cell characteristics.
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: Y J
authors.initials: YJ
authors.lastname: Yang
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: Z B
authors.initials: ZB
authors.lastname: Li
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: G R
authors.initials: GR
authors.lastname: Zhang
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: L J
authors.initials: LJ
authors.lastname: Wu
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: J Y
authors.initials: JY
authors.lastname: Yu
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: L J
authors.initials: LJ
authors.lastname: Hu
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: Y L
authors.initials: YL
authors.lastname: Zhou
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China.
authors.forename: H D
authors.initials: HD
authors.lastname: Wang
authors.affiliation: Hainan Province Nongken Sanya Hospital, Sanya, China liangdongnsh@126.com.
authors.forename: D
authors.initials: D
authors.lastname: Liang
mesh_terms: D000080924:Aldehyde Dehydrogenase 1 Family; D015703:Antigens, CD; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D050697:Retinal Dehydrogenase / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000071250:Snail Family Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D019076:Transgenes; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; C495270:CTNNB1 protein, human; D015820:Cadherins; D005353:Fibronectins; D007527:Isoenzymes; C000604407:SNAI1 protein, human; D000071250:Snail Family Transcription Factors; D014746:Vimentin; D051176:beta Catenin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000080924:Aldehyde Dehydrogenase 1 Family; C510223:ALDH1A1 protein, human; D050697:Retinal Dehydrogenase