ABSTRACT
Objective Tumor cells are able to support their malignant proliferation by changing metabolic models .Prostate cells rely much on lipid metabolism in which ATP-citrate lyase ( ACLY) plays a very important role .The aim of this research was to study the effects of downregulated ACLY on the cell proliferation , cycle distribution and apoptosis of androgen-independent prostate cancer cells DUl45. Methods DU145 cells were divided into two groups:the cells in experiment group were transfected with the small interfering RNA-mediated knockdown of ACLY , while the cells in control group were transfected with meaningless small interfering RNA.Cell counting Kit test ( CCK-8 ) was applied to detect the effects of the downregulation of ATP citrate lyase on the proliferation of DU145.Flow cytometry instrument was used to analyze the variation of cell cycle distribution and apoptosis rate between groups .Western blot was used to detect the change of intracellular Caspase-3 protein content. Results Western blot showed favorable effects of ACLY interference .Compared with control group , ACLY protein content significantly decreased in experiment group ( P0.05), while the percentage of G2 cells decreased and the percentage of S cells in-creased with most cell cycle blocking at G 0/G1 stage, which were of significant difference .Meanwhile the expression of apoptosis pro-tein Caspase-3 upregulated significantly . Conclusion ACLY is of vital significance to maintain the malignant proliferation of prostate cancer cells and its downregulation results in the inhibition of cell proliferation and the promotion of cell apoptosis .
ABSTRACT
Objective Intercellular adhesion molecule-1 (ICAM-1) plays an important role in mediating pulmonary infiltration of neutrophils .The aim of the study was to observe the expression of ICAM-1 and its potential regulators MK 2/HuR in pulmonary micro-vascular endothelial cells ( PMVEC ) in mice with acute respiratory distress syndrome ( ARDS) induced by lipopolysaccharide ( LPS) . Methods Ten 6-8 weeks old healthy C57BL/6 mice were randomly divided into an LPS and a control group of equal number , the former injected intraperitoneally with LPS diluted in 100 μL PBS while the latter with PBS only , both at 5 mg per kg of the body weight .At 24 hours after injection , all the mice were sacrificed .Real-time PCR was used to determine the mRNA expressions of HuR and ICAM-1 in the PMVECs, Western bolt employed to detect the protein expressions of MK2, HuR and ICAM-1, and flow cytometry adopted to measure the ICAM-1 expression on the surface of the PMVECs and pulmonary infiltration of neutrophils . Results The W/D ratio in the lung tissue of the mice was significantly lower in the LPS than in the control group (3.61 ±0.28 vs 6.16 ±0.40, P<0.05), while the rate of neutrophil infiltration markedly higher in the former than in the latter ([13.92 ±3.23]%vs [3.24 ±1.24]%, P<0.05).The mRNA and protein expressions of ICAM-1 in the PMVECs were significantly elevated in the LPS group as compared with that in the control (P<0.05), and so was the mRNA expression of HuR (P<0.05).No remarkable changes were observed in the expressions of total MK 2 and HuR proteins, but phosphorylated MK2 (p-MK2) and cytoplasmic HuR were increased in the LPS-stimulated mice. Conclusion Specific blockage or reduction of the HuR expression in PMVECs may lower the expression of ICAM-1, reduce neutrophil infiltration , and lessen pathophysiological changes in mice with ARDS .