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Chien S.-T.,Kaohsiung Armed Forces General Hospital | Chien S.-T.,Fooyin University | Shi M.-D.,Kaohsiung Veterans General Hospital Tainan Branch | Shi M.-D.,Chung Hwa University of Medical Technology | And 3 more authors.
Cancer Cell International | Year: 2015

Background: Galangin (3,5,7-trihydroxyflavone) is a flavonoid compound found in high concentration in lesser galangal. The objective of this study was to investigate the ability of galangin to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced the invasion and metastasis of HepG2 liver cancer cells. Results: First, using a cell-matrix adhesion assay, immunofluorescence assay, transwell-chamber invasion/migration assay, and wound healing assay, we observed that galangin exerted an inhibitory effect on TPA-induced cell adhesion, morphology/actin cytoskeleton arrangement, invasion and migration. Furthermore, the results of gelatin zymography and reverse transcriptase polymerase chain reaction (RT-PCR) assays showed that galangin reduced the TPA-induced enzyme activity of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in HepG2 cells; moreover, the messenger RNA level was downregulated. We also observed through a Western blotting assay that galangin strongly inhibited the TPA-induced protein expressions of protein kinase Cα (PKCα), protein kinase Cδ (PKCδ), phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), the phospho-inhibitor of kappaBα (phospho-IΚBα), c-Fos, c-Jun, and nuclear factor kappa B (NF-ΚB). Next, galangin dose-dependently inhibited the binding ability of NF-ΚB and activator protein 1 (AP-1) to MMP-2/MMP-9 promoters, respectively, resulting in the suppression of MMP-2/MMP-9 enzyme activity. Conclusions: The results revealed that galangin effectively inhibited the TPA-induced invasion and migration of HepG2 cells through a protein kinase C/extracellular signal-regulated kinase (PKC/ERK) pathway. Thus, galangin may have widespread applications in clinical therapy as an anti-metastatic medicament. © Chien et al.


Wang J.-L.,Kaohsiung Veterans General Hospital Tainan Branch | Chou C.-T.,Chang Gung Institute of Technology | Liang W.-Z.,Kaohsiung Veterans General Hospital | Kuo C.-C.,Tzu Hui Institute of Technology | And 5 more authors.
Toxicology Mechanisms and Methods | Year: 2016

The phenolic compound 2,5-dimethylphenol is a natural product. 2,5-Dimethylphenol has been shown to affect rat hepatic and pulmonary microsomal metabolism. However, the effect of 2,5-dimethylphenol on Ca2+ signaling and cyotoxicity has never been explored in any culture cells. This study explored the effect of 2,5-dimethylphenol on cytosolic free Ca2+ levels ([Ca2+]i) and cell viability in PC3 human prostate cancer cells. 2,5-Dimethylphenol at concentrations between 500 μM and 1000 μM evoked [Ca2+]i rises in a concentration-dependent manner. This Ca2+ signal was inhibited by approximately half by the removal of extracellular Ca2+. 2,5-Dimethylphenol-induced Ca2+ influx was confirmed by Mn2+-induced quench of fura-2 fluorescence. Pretreatment with the protein kinase C (PKC) inhibitor GF109203X, nifedipine or the store-operated Ca2+ entry inhibitors (econazole or SKF96365) inhibited 2,5-dimethylphenol-induced Ca2+ signal in Ca2+-containing medium by ∼30%. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin in Ca2+-free medium abolished 2,5-dimethylphenol-induced [Ca2+]i rises. Conversely, treatment with 2,5-dimethylphenol abolished thapsigargin-induced [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 reduced 2,5-dimethylphenol-evoked [Ca2+]i rises by ∼80%. 2,5-Dimethylphenol killed cells at concentrations of 350–1000 μM in a concentration-dependent fashion. Chelation of cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N, N, N′, N′-tetraacetic acid/AM (BAPTA/AM) did not prevent 2,5-dimethylphenol’s cytotoxicity. Together, in PC3 cells, 2,5-dimethylphenol induced [Ca2+]i rises that involved Ca2+ entry through PKC-regulated store-operated Ca2+ channels and PLC-dependent Ca2+ release from the endoplasmic reticulum. 2,5-Dimethylphenol induced cytotoxicity in a Ca2+-independent manner. © 2016 Informa UK Limited, trading as Taylor & Francis Group.


Lee H.-H.,St Joseph Hospital | Chou C.-T.,Chang Gung Institute of Technology | Liang W.-Z.,Kaohsiung Veterans General Hospital | Chen W.-C.,St Joseph Hospital | And 6 more authors.
Chinese Journal of Physiology | Year: 2016

This study explored the effect of deltamethrin, a pesticide, on intracellular free Ca2+ concentration ([Ca2+]i) in PC3 human prostate cancer cells. Deltamethrin at concentrations between 5 µM and 20 µM evoked [Ca2+]i rises in a concentration-dependent manner. This Ca2+ signal was inhibited by 22% by removal of extracellular Ca2+. Nifedipine, econazole, and SKF96365 also inhibited the Ca2+ signal. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) in Ca2+-free medium nearly abolished deltamethrin-induced [Ca2+]i rises. Treatment with deltamethrin also inhibited most of BHQ-induced [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 failed to alter deltamethrin-evoked [Ca2+]i rises. Deltamethrin killed cells at concentrations of 20-100 µM in a concentration-dependent fashion. Chelation of cytosolic Ca2+ with 1,2-bis (2-aminophenoxy) ethane-N, N, N’, N’-tetraacetic acid/acetoxymethyl ester (BAPTA/AM) did not prevent deltamethrin’s cytotoxicity. Together, in PC3 human prostate cancer cells, deltamethrin induced [Ca2+]i rises that involved Ca2+ entry through store-operated Ca2+ channels and PLC-independent Ca2+ release from the endoplasmic reticulum. Deltamethrin induced cytotoxicity in a Ca2+-independent manner. © 2016 by The Chinese Physiological Society and Airiti Press Inc.


Liu C.-P.,Yuans General Hospital | Chou C.-T.,Chang Gung University | Liang W.-Z.,Kaohsiung Veterans General Hospital | Chang H.-T.,Kaohsiung Veterans General Hospital Tainan Branch | And 5 more authors.
Journal of Receptors and Signal Transduction | Year: 2013

The effect of angiotensin II (Ang II) on cytosolic Ca2+ concentrations ([Ca2+]i) in MDCK renal tubular cells was explored. The Ca2+-sensitive fluorescent dye fura-2 was applied to measure [Ca2+]i. Ang II at concentrations of 5-40 M induced a [Ca 2+]i rise in a concentration-dependent manner. The response was reduced partly by removing Ca2+. Ang II evoked store-operated Ca 2+ entry that was inhibited by La3+ and Gd3+. In the absence of extracellular Ca2+, incubation with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) or thapsigargin abolished Ang II-induced Ca2+ release. Inhibition of phospholipase C with U73122 abolished Ang II-induced [Ca2+]i rise. Three Ang II analogues [(ASN1,VAL5)-Ang II acetate, (SAR1,THR8)-Ang II acetate, (VAL5)-Ang II acetate] failed to induce a [Ca2+]i rise. Together, in MDCK cells, Ang II induced a [Ca2+]i rise via Ca2+ entry through store-operated Ca2+ channels and phospholipase C-dependent Ca2+ release from the endoplasmic reticulum. Moreover, Ang II's amino acid sequence is important in its stimulatory effect on [Ca2+]i. © 2013 Informa Healthcare USA, Inc.


Wang S.-C.,Kaohsiung Veterans General Hospital Tainan Branch | Yu C.-L.,Kun Shan University
Aging and Mental Health | Year: 2015

Objectives: The purpose was to examine the effectiveness of music care on cognitive function, depression, and behavioral problems among elderly people with dementia in long-term care facilities in Taiwan. Methods: The study had a quasi-experimental, longitudinal research design and used two groups of subjects. Subjects were not randomly assigned to experimental group (n = 90) or comparison group (n = 56). Based on Bandura's social cognition theory, subjects in the experimental group received Kagayashiki music care (KMC) twice per week for 24 weeks. Subjects in the comparison group were provided with activities as usual. Results: Results found, using the control score of the Clifton Assessment Procedures for the Elderly Behavior Rating Scale (baseline) and time of attending KMC activities as a covariate, the two groups of subjects had statistically significant differences in the mini-mental state examination (MMSE). Results also showed that, using the control score of the Cornell Scale for Depression in Dementia (baseline) and MMSE (baseline) as a covariate, the two groups of subjects had statistically significant differences in the Clifton Assessment Procedures for the Elderly Behavior Rating Scale. Conclusion: These findings provide information for staff caregivers in long-term care facilities to develop a non-invasive care model for elderly people with dementia to deal with depression, anxiety, and behavioral problems. © 2015 Taylor & Francis

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