Study on diverse pathological characteristics of heart failure in different stages based on proteomics.

Heart failure is a process characterized by significant disturbance of protein turnover. To elucidate the alterations in cardiac protein expression during the various phases of heart failure and to understand the nature of the processes involved, we analysed the proteome in an established heart failure model at different time points to monitor thousands of different proteins simultaneously. Here, heart failure was induced by transverse aortic constriction (TAC) in KM mice. At 2, 4 and 12 weeks after operation, protein expression profiles were determined in sham-operated (controls) and TAC mice, using label-free quantitative proteomics, leading to identification and quantification of almost 4000 proteins. The results of the KEGG pathway enrichment analysis and GO function annotation revealed critical pathways associated with the transition from cardiac hypertrophy to heart failure, such as energy pathways and matrix reorganization. Our study suggests that in the pathophysiology of heart failure, alterations of protein groups related to cardiac energy substrate metabolism and cytoskeleton remodelling could play the more dominant roles for the signalling that eventually results in contractile dysfunction and heart failure.

Effects of mdig on proliferation and apoptosis of lung cancer cells.

Expression of mineral dust-induced gene (mdig) in lung cancer NCI-H1650 cells was detected to investigate the effects of mdig on proliferation and apoptosis of NCI-H1650 cells. NCI-H1650 lung cancer cells were cultured in vitro. The expression of mdig in NCI-H1650 cells was lowered using ribonucleic acid interference (RNAi) technique. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to detect the effects of mdig small interfering RNA (siRNA) on messenger RNA (mRNA) and the protein expression of mdig in lung cancer NCI-H1650 cells, respectively. The effect of mdig on the proliferation of NCI-H1650 cells was observed through 3-(4,5)-dimethylthiazol (-z-y1)-3,5-di-phenyl tetrazolium bromide (MTT) assay, and flow cytometry was used to detect the impact of mdig on cell cycle and apoptosis of NCI-H1650 cells. The influence of mdig on caspase-3 and poly (ADP-ribose) polymerase 1 (PARP1) proteins in NCI-H1650 cells were investigated via western blot analysis. The results of RT-qPCR and western blot analysis showed that mdig siRNA obviously inhibited the expression of mRNA and protein of mdig in NCI-H1650 cells. Results of the MTT assay showed mdig siRNA could significantly reduce the proliferation ability of NCI-H1650 cells. In addition cell cycle detection showed that mdig siRNA caused NCI-H1650 cell arrest at G1 phase. Apoptosis detection results indicated that mdig siRNA promoted apoptosis of NCI-H1650 cells. Western-blot analysis revealed that mdig siRNA upregulated the expression of cleaved caspase-3 and cleaved PARP1 proteins in NCI-H1650 cells. Mdig is highly expressed in lung cancer NCI-H1650 cells while mdig siRNA can inhibit proliferation of NCI-H1650 cells and accelerate apoptosis. The underlying mechanism may be related to inhibited cell cycle progression and upregulated expression of cleavage proteins (cleaved caspase-3 and cleaved PARP1).

MicroRNA-1246 inhibits cell invasion and epithelial mesenchymal transition process by targeting CXCR4 in lung cancer cells.

BACKGROUND: Recent studies have indicated that microRNAs (miRNAs) are closely related to lung cancer. However, the effects of miR-1246 on lung cancer are still elusive. In this study, we aimed to explore the molecular mechanisms of miR-1246 in lung cancer. MATERIALS AND METHODS: Using RT-qPCR assay, we analyzed the expression of miR-1246 in lung cancer cell lines and lung epithelial cell line. Using Cell Counting Kit-8 (CCK-8), flow cytometry, Transwell, RT-qPCR and western blot assays, we investigated cell viability, apoptosis, invasion and epithelial mesenchymal transition (EMT) process. Using luciferase reporter assay, we confirmed a target of miR-1246. Using western blot assay, we detected the protein mechanisms of Janus kinase (JAK)/signal transducer and activator of transcription (STAT) and phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) signal pathways. RESULTS: Our results showed that miR-1246 was down-regulated in lung cancer cell lines (A549, H1650 and H1299) compared to in lung epithelial cell line (16HBE14o). MiR-1246 overexpression remarkably inhibited cell invasion as well as up-regulated E-cadherin expression and down-regulated N-cadherin, Vimentin, ZEB1 and Snail expressions in A549 cells. Further studies have confirmed CXCR4 as a target gene of miR-1246, and CXCR4 silence significantly abolished the promotion effect of miR-1246 suppression on cell invasion and EMT process in A549 cells. Besides, miR-1246 blocked JAK/STAT and PI3K/AKT signal pathways by regulation of CXCR4. CONCLUSIONS: These results demonstrated that miR-1246 inhibited cell invasion and EMT process by targeting CXCR4 and blocking JAK/STAT and PI3K/AKT signal pathways in lung cancer cells.

A New Biomarkers Feature Pattern Consisting of TNF- α , IL-10, and IL-8 for Blood Stasis Syndrome with Myocardial Ischemia.

Objective. To explore new diagnostic patterns for syndromes to overcome the insufficiency of obtainable macrocharacteristics and specific biomarkers. Methods. Chinese miniswines were subjected to Ameroid constrictor, placed around the proximal left anterior descending branch. On the 4th week, macrocharacteristics, coronary angiography, echocardiography, and hemorheology indices were detected for diagnosis. IL-1, IL-6, IL-8, IL-10, TNF- α , and hsCRP in serum were detected, and Decision Tree was built. Results. According to current official-issued standard, model animals matched the diagnosis of blood stasis syndrome with myocardial ischemia based on findings, including >90% occlusion, attenuated left ventricular segmental motion, dark red or purple tongues, and higher blood viscosity. Significant decrease of IL-10 and increase of TNF- α were found in model animals. However, in the Decision Tree, besides IL-10 and TNF- α , IL-8 helped to increase the accuracy of classification to 86%. Conclusions. The Decision Tree building with TNF- α , IL-10, and IL-8 is helpful for the diagnosis of blood stasis syndrome in myocardial ischemia animals. What is more is that our data set up a new path to the differentiation of syndrome by feature patterns consisting of multiple biomarkers not only for animals but also for patients. We believe that it will contribute to the standardization and international application of syndromes.