ABSTRACT
We studied the effects and mechanisms of action of conophylline in different concentrations in the original in vitro model of myocardial fibrosis (treatment of cardiac fibroblasts isolated form the hearts of newborn rats with angiotensin II). Viability, collagen content, and expression of related protein in cardiac fibroblasts were assessed using the MTT-test, Sircol assay, and Western blotting, respectively. Conophylline markedly protected the cultured cells against the development of angiotensin II-induced fibrosis, which was seen from reduced viability of fibroblasts, decreased collagen content, and down-regulation of the expression of α-smooth muscle actin (α-SMA). Conophylline did not affect the TGF-ß pathway altered by angiotensin II, but markedly decreased the level of bone morphogenetic protein-4 (BMP4) enhanced by angiotensin II and BMP4 itself. Conophylline produced no effect on phosphorylation of α-SMA and Smad homologue-1/5/8, the classic BMP4 downstream pathway elements, but reduced the level of c-Jun N-terminal kinase (JNK) elevated by BMP4. Conophylline did not inhibit the development of myocardial fibrosis in the presence of JNK activator anisomycin. Thus, conophylline inhibited angiotensin II-provoked myocardial fibrosis via the BMP4/JNK pathway.
Subject(s)
Angiotensin II/pharmacology , Antifibrotic Agents/pharmacology , Bone Morphogenetic Protein 4/genetics , Fibroblasts/drug effects , MAP Kinase Kinase 4/genetics , Vinca Alkaloids/pharmacology , Animals , Animals, Newborn , Bone Morphogenetic Protein 4/antagonists & inhibitors , Bone Morphogenetic Protein 4/metabolism , Collagen/genetics , Collagen/metabolism , Endomyocardial Fibrosis/genetics , Endomyocardial Fibrosis/metabolism , Endomyocardial Fibrosis/pathology , Endomyocardial Fibrosis/prevention & control , Fibroblasts/metabolism , Fibroblasts/pathology , Gene Expression Regulation , MAP Kinase Kinase 4/antagonists & inhibitors , MAP Kinase Kinase 4/metabolism , Models, Biological , Myocardium/metabolism , Myocardium/pathology , Phosphorylation/drug effects , Primary Cell Culture , Rats , Rats, Wistar , Signal Transduction , Smad1 Protein/genetics , Smad1 Protein/metabolism , Smad5 Protein/genetics , Smad5 Protein/metabolism , Smad8 Protein/genetics , Smad8 Protein/metabolism , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism , p38 Mitogen-Activated Protein Kinases/genetics , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Liver cancer is a malignant cancer with great harmfulness. Fenofibrate is a peroxisome proliferation activated receptor (PPARα) agonist widely used in the treatment of dyslipidemia. Previous studies have shown that fenofibrate may promote cell proliferation, but the underlying mechanism has not been fully characterized. The aim of this study was to investigate the role of PPARα agonist fenofibrate in cell proliferation of SMMC-7721 cells compared with that of THLE-2 cells. SMMC-7721 and THLE-2 cells were treated with different concentrations of fenofibrate. Cell proliferation was analyzed by MTT, using flow cytometry for cell cycle analysis, and CyclinD1, Cyclin-dependent kinases2 (CDK2) and Proliferating Cell Nuclear Antigen (PCNA) were analyzed by Western blotting. RT-qPCR method was used to assess CDK2, CyclinD1 and PCNA mRNA levels. The results showed that 10-9-10-4 mol/L fenofibrate could induce cell growth and 10-4, 10-5, 10-6 mol/L fenofibrate could reduce the number of G0/G1 phase cells and increased in the number of cells in S and G2/M phase of cell cycle in SMMC-7721 cells. Furthermore, fenofibrate could significantly increase the expression of cell cycle related protein (CyclinD1, CDK2)and cell proliferation related proteins (PCNA). The use of PPARα inhibitor MT886 inhibited cell cycle progression and promote tumor cell apoptosis. But fenofibrate had no obvious effect on THLE-2 cells. These results revealed the effect of fenofibrate on the cell cycle of liver cancer cells, and provided a reasonable explanation for studying how fenofibrate promotes cell proliferation.
Subject(s)
Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Cell Cycle/drug effects , Cell Proliferation/drug effects , Fenofibrate/pharmacology , Liver Neoplasms/drug therapy , Tumor Cells, Cultured/drug effects , China , Humans , Hypolipidemic Agents/pharmacology , PPAR alpha/pharmacologyABSTRACT
OBJECTIVE: Oral squamous cell carcinoma (OSCC) is one of the most common head and neck tumors with high incidence and mortality. Long noncoding RNA bladder cancer-associated transcript 1 (lncRNA BLACAT1) was involved in several cancers development. However, the roles of BLACAT1 in OSCC have not been investigated. MATERIALS AND METHODS: The expressions of BLACAT1 and miR-142-5p in OSCC cells were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was evaluated by MTT assay. Cell migration and invasion were evaluated by transwell migration assay and transwell invasion assay, respectively. The protein levels of CyclinD1, p21, p27, MMP-2, MMP-9 and MMP-14 were detected by Western blot analysis. The interaction of BLACAT1 and miR-142-5p was verified by luciferase reporter assay. RESULTS: The expression of BLACAT1 was increased and the expression of miR-142-5p was decreased in OSCC cells. The knockdown of BLACAT1 suppressed the viability, migration and invasion of OSCC cells. miR-142-5p was identified as a target of BLACAT1 and BLACAT1 overexpression suppressed miR-142-5p expression. Furthermore, overexpression of miR-142-5p showed similar effects on OSCC cells viability, migration and invasion with BLACAT1 knockdown, and inhibition of miR-142-5p restored the effects of BLACAT1 knockdown OSCC cells viability, migration and invasion. CONCLUSIONS: LncRNA BLACAT1 knockdown suppressed the viability, migration and invasion of OSCC cells by sponging miR-142-5p, indicating that BLACAT1 might be a novel target for the treatment of OSCC.
