Epithelial requirement for in vitro proliferation and xenograft growth and metastasis of MDA-MB-468 human breast cancer cells: oncogenic rather than tumor-suppressive role of E-cadherin

Autor: H. J. Hugo, N. P. A. D. Gunasinghe, B. G. Hollier, T. Tanaka, T. Blick, A. Toh, P. Hill, C. Gilles, M. Waltham, E. W. Thompson
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Breast Cancer Research, Vol 19, Iss 1, Pp 1-25 (2017)
Druh dokumentu: article
ISSN: 1465-542X
DOI: 10.1186/s13058-017-0880-z
Popis: Abstract Background Epithelial-to-mesenchymal transition (EMT) is associated with downregulated E-cadherin and frequently with decreased proliferation. Proliferation may be restored in secondary metastases by mesenchymal-to-epithelial transition (MET). We tested whether E-cadherin maintains epithelial proliferation in MDA-MB-468 breast cancer cells, facilitating metastatic colonization in severe combined immunodeficiency (SCID) mice. Methods EMT/MET markers were assessed in xenograft tumors by immunohistochemistry. Stable E-cadherin manipulation was effected by transfection and verified by Western blotting, immunocytochemistry, and quantitative polymerase chain reaction (qPCR). Effects of E-cadherin manipulation on proliferation and chemomigration were assessed in vitro by performing sulforhodamine B assays and Transwell assays, respectively. Invasion was assessed by Matrigel outgrowth; growth in vivo was assessed in SCID mice; and EMT status was assessed by qPCR. Hypoxic response of E-cadherin knockdown cell lines was assessed by qPCR after hypoxic culture. Repeated measures analysis of variance (ANOVA), one- and two-way ANOVA with posttests, and paired Student’s t tests were performed to determine significance (p
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