title: The effect of TGF-beta-induced epithelial-mesenchymal transition on the expression of intracellular calcium-handling proteins in T47D and MCF-7 human breast cancer cells.
abstract: The contribution of Ca(2+) in TGF-β-induced EMT is poorly understood. We aimed to confirm the effect of TGF-β on the gene expression of intracellular calcium-handling proteins and to investigate the potential underlying mechanisms in TGF-β-induced EMT. T47D and MCF-7 cells were cultured in vitro and treated with TGF-β. The mRNA expression of EMT marker genes and intracellular calcium-handling proteins were quantified by qRT-PCR. qRT-PCR and Western blot analysis results verified the changes of EMT marker gene expression. Furthermore, we found that TGF-β induced cell morphological changes significantly with an increase of cell surface area and cell length. These results indicated that TGF-β induced EMT. The mRNA expression levels of SPCA1, SPCA2 and MCU were not influenced by TGF-β treatment, while NCX1 expression was decreased in T47D cells. In addition, the mRNA levels of SERCAs and IP3Rs were significantly changed due to TGF-β-induced EMT. The TGF-β-treated T47D cells exhibited markedly greater response to ATP than the control cells, and the descent velocity of cytosolic calcium concentration was faster in TGF-β-treated cells than in control cells. This is the first report to demonstrate that TGF-β-induced EMT in human breast cancer cells is associated with alterations in endoplasmic reticulum calcium homeostasis.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China; State Key Laboratory of Biomembrane and Membrane Biotechnology, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Shah H A
authors.initials: SH
authors.lastname: Mahdi
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Huanyi
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Breast Cancer Center, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University Cancer Hospital & Institute, Beijing 100142, China.
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China. Electronic address: rqfeng@pku.edu.cn.
authors.forename: Renqing
authors.initials: R
authors.lastname: Feng
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D061986:MCF-7 Cells; D015388:Organelles / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
chemical_list: D015220:Calcium Channels; D016212:Transforming Growth Factor beta; D012313:RNA; D002118:Calcium
keywords: Breast cancer; Calcium channel; Calcium pump; EMT; NCX; TGF-β