Abstract: The calcium‐sensing receptor (CaR) is a G protein‐coupled receptor. The CaR stimulation elicits phospholipase C‐mediated inositol triphosphate formation, leading to an elevation in the level of intracellular calcium released from endoplasmic reticulum (ER). Depletion of ER Ca 2+ leads to ER stress, which is thought to induce apoptosis. Intracellular calcium overload‐induced apoptosis in cardiac myocytes during hypoxia–reoxygenation (H/Re) has been demonstrated. However, the links between CaR, ER stress and apoptosis during H/Re are unclear. This study hypothesized that the CaR could induce apoptosis in neonatal rat cardiomyocytes during H/Re via the ER stress pathway. Neonatal rat cardiomyocytes were subjected to 3 hr of hypoxia, followed by 6 hr of reoxygenation. CaR expression was elevated and the number of apoptotic cells was significantly increased, as shown by transferase‐mediated dUTP nick end‐labelling, with exposure to CaCl 2 , a CaR activator, during H/Re. The intracellular calcium concentration was significantly elevated and the Ca 2+ concentration in the ER was dramatically decreased during H/Re with CaCl 2 ; both intracellular and ER calcium concentrations were detected by laser confocal microscopy. Expression of GRP78 (glucose‐regulated protein 78), the cleavage products of ATF6 (activating transcription factor 6), phospho‐PERK [pancreatic ER kinase (PKR)‐like ER kinase], the activated fragments of caspase‐12, and phospho‐JNK (c‐Jun NH 2 ‐terminal kinase) were increased following exposure to CaCl 2 during H/Re. Our results confirmed that the activated CaR can induce cardiomyocyte apoptosis via ER stress‐associated apoptotic pathways during H/Re.
Triple-negative breast cancer (TNBC) is highly invasive and metastatic, which is in urgent need of transformative therapeutics. Tubeimu (TBM), the rhizome of Bolbostemma paniculatum (Maxim.) Franquet, is one of the Chinese medicinal herbs used for breast diseases since the ancient times. The present study evaluated the efficacy, especially the anti-metastatic effects of the dichloromethane extract of Tubeimu (ETBM) on TNBC orthotopic mouse models and cell lines.We applied real-time imaging on florescent orthotopic TNBC mice model and tested cell migration and invasion abilities with MDA-MB-231 cell line. Digital gene expression sequencing was performed and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis applied to explore the pathways influenced by ETBM. Moreover, quantitative real-time polymerase chain reactions (qRT-PCR) and Western blot were delivered to confirm the gene expression changes.ETBM exhibited noticeable control on tumor metastasis and growth of TNBC tumors with no obvious toxicity. In compliance with this, it also showed inhibition of cell migration and invasion in vitro. Its impact on the changed biological behavior in TNBC may be a result of decreased expression of integrin β1 (ITGβ1), integrin β8 (ITGβ8) and Rho GTPase activating protein 5 (ARHGAP5), which disabled the focal adhesion pathway and caused change in cell morphology.This study reveals that ETBM has anti-metastatic effects on MDA-MB-231-GFP tumor and may lead to a new therapeutic agent for the integrative treatment of highly invasive TNBC.
Abstract Hepatocellular carcinoma (HCC) is the major type of primary liver cancer. Genomic studies have revealed that HCC is a heterogeneous disease with multiple subtypes. BRG1, encoded by the SMARCA4 gene, is a key component of SWI/SNF chromatin-remodeling complexes. Based on TCGA studies, somatic mutations of SMARCA4 occur in ~3% of human HCC samples. Additional studies suggest that BRG1 is overexpressed in human HCC specimens and may promote HCC growth and invasion. However, the precise functional roles of BRG1 in HCC remain poorly delineated. Here, we analyzed BRG1 in human HCC samples as well as in mouse models. We found that BRG1 is overexpressed in most of human HCC samples, especially in those associated with poorer prognosis. BRG1 expression levels positively correlate with cell cycle and negatively with metabolic pathways in the Cancer Genome Atlas (TCGA) human HCC data set. In a murine HCC model induced by c-MYC overexpression, ablation of the Brg1 gene completely repressed HCC formation. In striking contrast, however, we discovered that concomitant deletion of Brg1 and overexpression of c-Met or mutant NRas (NRAS V12 ) triggered HCC formation in mice. Altogether, the present data indicate that BRG1 possesses both oncogenic and tumor-suppressing roles depending on the oncogenic stimuli during hepatocarcinogenesis.
The aim of this study was to evaluate the effects of a collagen/hyaluronic acid coating without or with incorporated heterodimeric bone morphogenetic protein 2/7 (BMP2/7) on in-vitro osteoblastic differentiation on titanium discs. The multilayer collagen/hyaluronic acid coatings without or without incorporated BMP2/7 were deposited on titanium discs via a layer-by-layer technique. The effects of the coatings were evaluated by assessing the alkaline phosphatase (ALP) activity (an early osteoblastic differentiation marker) and the osteocalcin expression (a late osteoblastic differentiation marker). The expression levels of the osteoblastic genes, such as alkaline phosphatase 2 (AKP2) and osteocalcin (OC) were detected using real-time RT-PCR. ALP activity and OC expression were significantly increased when cells were cultured with collagen/hyaluronic acid+BMP2/7 heterodimer (p<0.05). The same result was found in cells with the expression of a BMP2/7 fusion gene, OC and AKP2. These results indicated that collagen/hyaluronic acid+BMP2/7 heterodimer-coated discs might have the potential to greatly enhance osseointegration than a either BMP2 or BMP7 solution or a mixture of BMP2 and BMP7 BMP2/7.
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