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.

The contribution of Ca(2+) in TGF-ß-induced EMT is poorly understood. We aimed to confirm the effect of TGF-ß on the gene expression of intracellular calcium-handling proteins and to investigate the potential underlying mechanisms in TGF-ß-induced EMT. T47D and MCF-7 cells were cultured in vitro and treated with TGF-ß. 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-ß induced cell morphological changes significantly with an increase of cell surface area and cell length. These results indicated that TGF-ß induced EMT. The mRNA expression levels of SPCA1, SPCA2 and MCU were not influenced by TGF-ß treatment, while NCX1 expression was decreased in T47D cells. In addition, the mRNA levels of SERCAs and IP3Rs were significantly changed due to TGF-ß-induced EMT. The TGF-ß-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-ß-treated cells than in control cells. This is the first report to demonstrate that TGF-ß-induced EMT in human breast cancer cells is associated with alterations in endoplasmic reticulum calcium homeostasis.

Physiological characterization of the cold-shock-induced humoral factor for wing color-pattern changes in butterflies.

Butterfly wing color patterns can be modified by the application of temperature shock to pupae immediately after pupation, which has been attributed to a cold-shock-induced humoral factor called cold-shock hormone (CSH). Here, we physiologically characterized CSH and pharmacological action of tungstate, using a nymphalid butterfly Junonia orithya. We first showed that the precise patterns of modification were dependent on the time-point of the cold-shock treatment after pupation, and confirmed that the modification properties induced in a cold-shocked pupa were able to be transferred to another pupa in a parabiosis experiment. Cold-shock application after removal of the head and prothorax together still produced modified wings, excluding major involvement of the brain-retrocerebral neuroendocrine complex. Furthermore, tungstate injection induced modifications even in individuals whose head and prothorax were removed. Importantly, transplantation of tracheae isolated from cold-shocked pupae induced modifications in the recipient wings. We identified a chemical peak in hemolymph of the cold-shocked individuals using HPLC, which corresponded to dopamine, and demonstrated that dopamine and its related biogenic amines have ability to induce small color-pattern changes. Taken together, the present study suggests that CSH is likely to be secreted from trachea-associated endocrine cells upon cold-shock treatment and that tungstate may change color patterns via its direct action on wings.