title: Notch4+ cancer stem-like cells promote the metastatic and invasive ability of melanoma.
abstract: Sphere formation in conditioned serum-free culture medium supplemented with epidermal growth factor and basic fibroblast growth factor (tumorospheres) is considered useful for the enrichment of cancer stem-like cells, also known as tumor-initiating cells. We used a gene expression microarray to investigate the gene expression profile of melanoma cancer stem-like cells (MCSLCs). The results showed that MCSLCs highly expressed the following Notch signaling pathway molecules: Notch3 (NM_008716), Notch4 (NM_010929), Dtx4 (NM_172442), and JAG2 (NM_010588). Immunofluorescence staining showed tumorosphere cells highly expressed Notch4. Notch4(high) B16F10 cells were isolated by FACS, and Western blotting showed that high Notch4 expression is related to the expression of epithelial-mesenchymal transition (EMT)-associated proteins. Reduced invasive and migratory properties concomitant with the downregulation of the EMT markers Twist1, vimentin, and VE-cadherin and the overexpression of E-cadherin was observed in human melanoma A375 and MUM-2B cells. In these cells, Notch4 was also downregulated, both by Notch4 gene knockdown and by application of the γ-secretase inhibitor, DAPT. Mechanistically, the re-overexpression of Twist1 by the transfection of cells with a Twist1 expression plasmid led to an increase in VE-cadherin expression and a decrease in E-cadherin expression. Immunohistochemical analysis of 120 human melanoma tissues revealed a significant correlation between the high expression of Notch4 and the metastasis of melanoma. Taken together, our findings indicate that Notch4+ MCSLCs trigger EMT and promote the metastasis of melanoma cells.
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.
authors.forename: Xian
authors.initials: X
authors.lastname: Lin
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.|Department of Pathology, Cancer Hospital of Tianjin Medical University, Tianjin, China.|Department of Pathology, General Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Baocun
authors.initials: B
authors.lastname: Sun
authors.affiliation: Department of Surgery, Stomatological Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Dongwang
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.|Department of Pathology, General Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Xiulan
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Surgery, Tianjin Hospital of ITCWM Nankai Hospital, Tianjin, China.
authors.forename: Ran
authors.initials: R
authors.lastname: Sun
authors.affiliation: Department of Pathology, Cancer Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.
authors.forename: Danfang
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.
authors.forename: Xueyi
authors.initials: X
authors.lastname: Dong
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.|Department of Pathology, General Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Gu
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.
authors.forename: Yanlei
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pathology, Tianjin Medical University, Tianjin, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
mesh_terms: D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D009361:Neoplasm Invasiveness / Q000235:genetics; D009362:Neoplasm Metastasis / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology*; D009687:Nuclear Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000074562:Receptor, Notch4; D051880:Receptors, Notch / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D051879:Twist-Related Protein 1 / Q000378:metabolism
chemical_list: D015820:Cadherins; C106075:NOTCH4 protein, human; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D000074562:Receptor, Notch4; D051880:Receptors, Notch; C103173:TWIST1 protein, human; D051879:Twist-Related Protein 1
keywords: Cancer stem cell; Notch4 protein; Twist1; epithelial-mesenchymal transition; melanoma