Abstract The paper aimed to investigate the effects of up-regulation and down-regulation of Mir-299-3p on the proliferation, migration, invasion and apoptosis of lung adenocarcinoma cell line H1299, and to preliminarily explore the mechanism of Mir-299-3p regulating the biological behavior of lung adenocarcinoma cells. Mir-299-3p inhibitor and Mir-299-3p mimics were transfected into lung adenocarcinoma cell line H1299, and the transfection efficiency was measured by fluorescence microscopy. The successfully transfected lung adenocarcinoma cells were detected of proliferation, migration, invasion, and apoptosis. Mir-299-3p inhibitor and Mir-299-3p mimics were used to down-regulate and up-regulate Mir-299-3p respectively. Western blot was employed to detect the expressions of STAT3, p-STAT3, caspase3, Bcl-2, Bax and MMP-9 proteins in lung adenocarcinoma cells. Small interfering RNA technology was applied to construct a cell line with low expression of PTPRD. WB was used to measure the transfection efficiency to determine whether the down-regulation of PTPRD can partially or completely reverse the proliferation, migration, invasion, and apoptosis of lung adenocarcinoma cells after the down-regulation of Mir-299-3p. It was also determined whether the down-regulation of PTPRD can partially or completely reverse the effect of Mir-299-3p down-regulation on the expressions of p-STAT3, MMP-9, Cleaved-caspase3, Bcl-2 and Bax proteins in lung adenocarcinoma cells. Down-regulation of Mir-299-3p can inhibit the proliferation and invasion of lung adenocarcinoma cell H1299, promote cell apoptosis, inhibit the expression of p-STAT3 and MMP-9, promote the expressions of apoptosis-inducing proteins such as Bax and Cleaved-caspase3, and also inhibit the expression of apoptosis inhibitor protein Bcl-2. The up-regulation of Mir-299-3p worked in the opposite way. PTPRD is the direct target gene of Mir-299-3p. Mir-299-3p may directly target PTPRD to regulate the expressions of p-STAT3, MMP-9, Cleaved-caspase3, Bcl-2 and Bax proteins to affect the biological behavior of lung adenocarcinoma cells.

OBJECTIVES: Hypoxia leads to endothelial cell inflammation, apoptosis, and damage, which plays an important role in the complications associated with ischemic cardiovascular disease. As an oxidoreductase, p66Shc plays an important role in the regulation of reactive oxygen species (ROS) production and apoptosis. Ketamine is widely used in clinics. This study was designed to assess the potential protective effect of ketamine against hypoxia-induced injury in human umbilical vein endothelial cells (HUVECs). Moreover, we explored the potential mechanism by which ketamine protected against hypoxia-induced endothelial injury. METHODS: The protective effects of ketamine against hypoxia-induced injury was assessed using cell viability and adhesion assays, quantitative polymerase chain reaction, and western blotting. RESULTS: Our data showed that hypoxia reduced HUVEC viability, increased the adhesion between HUVECs and monocytes, and upregulated the expression of endothelial adhesion molecules at the protein and mRNA levels. Moreover, hypoxia increased ROS accumulation and upregulated p66Shc expression. Furthermore, hypoxia downregulated sirt1 expression in HUVECs. Alternatively, ketamine was shown to reverse the hypoxia-mediated reduction of cell viability and increase in the adhesion between HUVECs and monocytes, ameliorate hypoxia-induced ROS accumulation, and suppress p66Shc expression. Moreover, EX527, a sirt1 inhibitor, reversed the protective effects of ketamine against the hypoxia-mediated reduction of cell viability and increase in adhesion between HUVECs and monocytes. CONCLUSION: Ketamine reduces hypoxia-induced p66Shc expression and attenuates ROS accumulation via upregulating sirt1 in HUVECs, thus attenuating hypoxia-induced endothelial cell inflammation and apoptosis.

Abstract Purpose: To investigate the mechanisms by which PD98059 and LY294002 interfere with the abnormal deposition of extracellular matrix regulated by connective tissue growth factor (CTGF) of rat pulmonary artery smooth muscle cells (PASMCs). Methods: Rat PASMCs were cultured and separated into a control group. Real-time fluorescence quantitative PCR was performed to detect the expression of collagen III and fibronectin mRNA. Immunohistochemistry and western blot analyses were performed to detect the expression of collagen III protein. Results: The expression of collagen III and fibronectin mRNA was greater in PASMCs stimulated with CTGF for 48 h, than in the control group. After 72h of stimulation, the expression of collagen III protein in the PASMCs was greater than in the control. The equivalent gene and protein expression of the CPL group were much more significant. Conclusions: CTGF can stimulate the gene expression of collagen III and fibronectin in PASMCs, which may be one of the factors that promote pulmonary vascular remodeling (PVR) under the conditions of pulmonary arterial hypertension (PAH). PD98059 and LY294002 can inhibit the ERK1/2 and PI3K/PKB signaling pathways, respectively, thus interfering with the biological effects of CTGF. This may be a new way to reduce PAH-PVR.