Regulation of valproic acid induced EMT by AKT/GSK3ß/ß-catenin signaling pathway in triple negative breast cancer.

Valproic acid (VPA) is a selective histone deacetylation (HDAC) inhibitor and exerts anti-cancer properties in different types of cancer. The epithelial-to-mesenchymal transition (EMT) mediating by different signaling cascade can be a potential target in aggressive human cancers. Therefore, we aimed to clarified the unravel relationship between AKT/GSK3ß/ß-catenin signalling pathway and VPA-induced EMT in triple negative breast cancer (TNBC). The cytotoxicity of VPA in MDA-MB-231 TNBC and MCF-10A control cells was evaluated. Alterations in the expression levels of Snail, E-cadherin, AKT, GSK3ß, ß-catenin were analyzed by RT-PCR. Additionally, Annexin V, cell cycle and wound healing assays were performed. Our results showed that VPA remarkably inhibited the growth of TNBC cell and triggered apoptotic cell death through G0/G1 arrest. Furthermore, VPA increased cell migration and activated the EMT process through significantly increasing Snail expression and in turn downregulation of E-cadherin and GKS3ß levels. However, the level of AKT and ß-catenin was reduced after treatment of VPA. Our data showed that VPA induced EMT process and cell migration in TNBC cells. However, AKT/GSK3ß/ß-catenin signaling pathway did not mediate EMT activation.

Effects of 1,25-Dihydroxy vitamin D3 on TNF-α induced inflammation in human chondrocytes and SW1353 cells: a possible role for toll-like receptors.

The purpose of this study is to evaluate anti-inflammatory and chondro-protective effects of 1,25(OH)2D3 in human chondrocytes and SW1353 cells via investigating expressions of MMPs, TIMPs, VDR, and intracellular signalling pathway mediators such as TLR-2 and -4. The HC and SW1353 cells were treated with 1,25(OH)2D3 at 10, 100, and 1000 nM concentrations in the absence/presence of TNF-α (20 ng/mL) for 48 h. The mRNA expressions of MMP-1, -2, -3, -9, and -13, TIMP-1 and -2, VDR, TLR-2 and -4 in HC and SW1353 cells were detected by qPCR after treatments. The cytotoxicity and cell proliferation analyses were assessed by LDH and WST-1 assay, respectively. Protein levels of MMPs, TIMPs, and VDR were analysed by immunocytochemistry and ELISA methods. TNF-α markedly increased cytotoxicity for 24, 48, 72 h (p < 0.05) and vitamin D treatment was shown to diminish the cytotoxic effect of TNF-α. Cell proliferations increased by Vitamin D in a dose-dependent manner. mRNA expressions of MMP-1, -2, -3, -9, and -13, TLR-2 and -4 genes decreased with 1,25(OH)2D3 treatment (p < 0.05). VDR, TIMP-1 and -2 levels elevated after TNF-α exposure compared with the control group in HC cells (p < 0.05). Protein expression levels were determined using Western blotting, ELISA and immunocytochemistry. 1,25(OH)2D3 via binding to VDR, reversed the effects of TNF-α by inhibiting TLR-2 and 4. Decreased levels of VDR, TIMP-1 and -2 after TNF-α treatment were elevated by 1,25(OH)2D3 proportional with increasing 1,25(OH)2D3 doses. 1,25(OH)2D3 and TNF-α co-treatment decreased MMP-1, -2, -3, -9, and -13 levels were after TNF-α exposure.

Hemodilution is not the only reason of difference: Comparison of fasting and non-fasting lipoproteins in paired samples.

INTRODUCTION: Low-density lipoprotein cholesterol (LDL) is an important risk factor for cardiovascular disease (CVD) and generally measured after 8-12 h fasting. However, some recent studies have pointed that non-fasting lipoproteins, especially LDL concentrations, are better indicators for demonstrating CVD risk and atherosclerosis. They asserted that nutrition is a negligible factor on changes in lipoprotein concentrations and claimed this difference as a result of hemodilution effect, caused from fluid intake and can be eliminated by applying some adjustments. We aimed to compare the fasting and non-fasting LDL values of the same individuals and discuss whether non-fasting and fasting LDL results can be used in place of each other, directly or after applying hemodilution correction models. MATERIAL AND METHODS: Fasting and non-fasting blood samples of 248 apparently healthy participants were collected. Lipid panel tests, albumin and hemoglobin levels were studied in each sample. Results were evaluated in seven different models which were recommended to correct the hemodilution effect on fasting and non-fasting lipid concentrations of the same individual. Concordance of fasting and non-fasting risk group of the individual were calculated according to the National Cholesterol Education Program classification. RESULTS: Fasting and non-fasting LDL and non-high density lipoprotein cholesterol (non-HDL) concentrations were significantly different in every model (p < 0.001). Concordance results of fasting and non-fasting LDL and non-HDL risk groups were 63.8% and 77.9% respectively. CONCLUSIONS: Our results demonstrated that fasting and non-fasting LDL and non-HDL concentrations could not be used in place of each other even when the results were adjusted for elimination of the hemodilution effect.