Promotion of Metastasis-associated Gene Expression in Survived PANC-1 Cells Following Trichostatin A Treatment.

OBJECTIVES: Histone deacetylase inhibitors represent a promising class of potential anticancer agents for the treatment of human malignancies. In this study, the effects of trichostatin A (TSA) on apoptosis, metastasis-associated gene expression, and activation of the Notch pathway in human pancreatic cancer cell lines were investigated. METHODS: After treatment with TSA, cell viability and apoptosis were evaluated using the MTT [3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide] assay, Hoechst 33258 staining, and flow cytometry. Moreover, RT-PCR and western blot analyses were performed to measure the expression levels of apoptosis-associated genes (Bcl-2, Bax, and caspase-3), metastasis-associated genes (E-cadherin, vimentin, and matrix metalloproteinases), and Notch pathway activation (Notch intracellular domain, NICD). The levels of matrix metalloproteinase 2 and NICD were also semi-quantified by immunoassay. RESULTS: Following treatment with TSA for 24 h, PANC-1, SW1990, and MIATACA-2 cells exhibited cell death. The MTT assay revealed that TSA significantly decreased cell viability in a dose-dependent manner in PANC-1 cells. The Hoechst 33258 staining and flow cytometry results evidenced a significant increase in PANC-1 cell apoptosis following TSA treatment. The expression levels of Bax and caspase-3 were increased significantly, whereas Bcl-2 was down-regulated after TSA treatment. In the PANC-1 cells that survived after TSA treatment, the expression levels of vimentin, E-cadherin, and MMP genes were altered by the promotion of potential metastasis and increased expression of NICD. CONCLUSIONS: TSA can induce apoptosis of pancreatic cancer cells. In addition, the up-regulation of metastasis-related genes and the activation of the Notch pathway in the survived PANC-1 cells may be associated with a too-low level of TSA or resistance to TSA.

Lipoxin A4 attenuation of endothelial inflammation response mimicking pancreatitis-induced lung injury.

Lipoxins (LXs) and their analogues are known to display potent anti-inflammatory actions. Previously, we reported that lipoxin A4 (LXA4) possessed powerful anti-inflammatory properties in acute pancreatitis in rats and that it may ameliorate the concomitant acute lung injury by reducing cytokine generation and inhibiting neutrophil activation. Considering that the vascular endothelium plays an important role during adherence, migration and activation of leukocytes, the present study was designed to investigate the effects of LXA4 on the inflammatory response induced by tumor necrosis factor α (TNF-α) in human pulmonary microvascular endothelial cells (HPMECs) and explore the potential mechanisms involved in these processes. We found that LXA4 markedly down-regulated the expression of monocyte chemotactic protein-1 (MCP-1), E-selectin, and interleukin-6 (IL-6) mRNA, as well as intercellular adhesion molecule-1 (ICAM-1) in TNF-α-exposed HPMECs. Moreover, LXA4 inhibited the phosphorylation and nuclear translocation of nuclear factor-κB/p65 (NF-κB/p65) and phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) in HPMECs following TNF-α stimulation. Heme oxygenase-1 (HO-1), a cytoprotective enzyme, was up-regulated by LXA4 in both non- and TNF-α-stimulated HPMECs. In conclusion, the protective effects of LXA4 to ALI may be executed through inhibition inflammation pathways of NF-κB and p38 MAPK and up-regulation of cytoprotective HO-1.

The meta-analysis of the association of PPARG P12A, C161T polymorphism and coronary heart disease.

OBJECTIVE: Two common variations of peroxisome proliferator-activated receptor γ (PPARG), P12A (Pro12Ala, rs1801282), and C161T (His447His, rs3856806), are thought to have an effect on susceptibility to coronary heart disease (CHD), but the results are inconsistent. Therefore, a meta-analysis of published studies was performed. METHODS: The electronic databases, PubMed, Embase, Web of Science, and CNKI (China National Knowledge Infrastructure) were searched for studies to include in the present meta-analysis (last search was updated on 30 Aug. 2011). Twenty studies testing the association between PPARG gene polymorphisms and CHD were examined: 12 studies of P12A; 8 studies of C161T. Overall and ethnicity-specific summary odds ratios and corresponding 95 % confidence intervals for CHD associated with these polymorphisms were estimated using fixed- and random-effects models. Heterogeneity and publication bias were evaluated. A total of 20 studies including 5,795 cases and 9,069 controls were included in this meta-analysis. RESULTS: No significant associations were found in carriers of the rare Ala allele of the P12A polymorphism versus the common Pro/Pro genotype among the studies with both of the fixed-effect and random-effect model. In the subgroup analyses by ethnicity, source of control and type of study, no significant risks were found. For PPARG C161T, carriers of the T variant of C161T polymorphism were associated with an increased risk of CHD (OR = 1.182, 95 % CI: 1.023-1.341, P(heterogeneity) = 0.002), and in the stratified analysis by ethnicity and source of controls, the contrast of CT + TT vs. CC all produced significant association in Asian and hospital-based controls (OR = 1.276, 95 % CI: 1.084-1.468, P(heterogeneity) = 0.055; OR = 1.164, 95 % CI: 1.001-1.326, P(heterogeneity) = 0.002),when the fixed-effect model was used. But they were all insignificant with the random-effect model. CONCLUSION: This meta-analysis suggests that the PPARG C161T polymorphism marginally contributes to increased susceptibility to CHD and marginally increased association between PPARG H477H polymorphism and CHD also appeared in Asian and hospital-based controls. But PPARG P12A polymorphism is not associated with CHD risk.

Glutathione S-transferases T1 null genotype is associated with susceptibility to aristolochic acid nephropathy.

OBJECTIVES: This study aims to determine whether six polymorphisms of the genes involved in drug metabolism are associated with susceptibility to the development and progression of aristolochic acid nephropathy (AAN). METHODS: In the study, 91 aristolochic acid nephropathy (AAN) cases and 152 healthy controls of Chinese Han population were examined. Six common polymorphisms of genes, including multidrug resistance gene 1 (MDR1), cytochrome P450 (CYP1A1), NAD(P)H quinone oxidoreductase 1 (NQO1), glutathione S-transferase (GST) T1 and M1, were determined. Associations between their genotypes with AAN risk were calculated using an unconditional logistic regression model. RESULTS: Among the six candidate polymorphisms, only the distribution frequency of GSTT1 null genotype was significantly higher among AAN cases compared with controls (P = 0.041, 62.6% vs. 48.7%) and was associated with a 1.7-fold increased risk (OR = 1.728, 95%CI: 1.013-2.948, P = 0.045) of developing AAN, after adjustment for age and gender. The stratified analysis further showed that the GSTT1 null genotype was dominant in slow progressive AAN patients (OR = 2.497, 95%CI: 1.028-6.064, P = 0.043). The GSTM1 genotypes were not shown to influence the development of AAN. CONCLUSION: This study supports the hypothesis that polymorphisms related to drug metabolism such as GSTT1 may be an important factor influencing the development of AAN in the Chinese Han population exposed to AA.