Troglitazone Inhibits Matrix Metalloproteinase-9 Expression and Invasion of Breast Cancer Cell through a Peroxisome Proliferator-Activated Receptor γ-Dependent Mechanism / 한국유방암학회지

PURPOSE: Peroxisome proliferator-activated receptor γ (PPARγ) is involved in the pathology of numerous diseases including atherosclerosis, diabetes, obesity, and cancer. Matrix metalloproteinases (MMPs) play a significant role in tissue remodeling related to various processes such as morphogenesis, angiogenesis, tissue repair, invasion, and metastasis. We investigated the effects of PPARγ on MMP expression and invasion in breast cancer cells. METHODS: MCF-7 cells were cultured and then cell viability was monitored in an MTT assay. Western blotting, gelatin zymography, real-time polymerase chain reaction, and luciferase assays were performed to investigate the effect of the synthetic PPARγ ligand troglitazone on MMP expression. Transcription factor DNA binding was analyzed by electrophoretic mobility shift assay. A Matrigel invasion assay was used to assess the effects of troglitazone on MCF-7 cells. RESULTS: Troglitazone did not affect MCF-7 cell viability. 12-O-tetradecanoylphorbol-13-acetate (TPA) induced MMP-9 expression and invasion in MCF-7 cell. However, these effects were decreased by troglitazone. TPA increased nuclear factor κB and activator protein-1 DNA binding, while troglitazone inhibited these effects. The selective PPARγ antagonist GW9662 reversed MMP-9 inhibition by troglitazone in TPA-treated MCF-7 cells. CONCLUSION: Troglitazone inhibited nuclear factor κB and activator protein-1-mediated MMP-9 expression and invasion of MCF-7 cells through a PPARγ-dependent mechanism.

Influencing Factors Affecting Successful Organ Donation and Families' Consent to Donate after Drain Death / 대한이식학회지

BACKGROUND: This study was conducted to identify the factors influencing successful organ donation and families' consent to donate after brain death. METHODS: Medical records and data regarding organ donation counseling with the families of 107 brain-dead potential donors between September 2012 and March 2016 at a single tertiary medical center were retrospectively reviewed. RESULTS: The final consent rate was 57.9% (62/107), and successful donation was performed in 40.2% (43/107) of cases. Univariate and multivariate analyses revealed that the independent factors associated with successful organ donation were age <60 years (odds ratio [OR], 3.384; 95% confidence interval [CI], 1.350 to 8.484; P=0.009), systolic blood pressure ≥90 mmHg (OR, 6.261; 95% CI, 1.418 to 27.653; P=0.015), and serum sodium level ≥150 mEq (OR, 4.215; 95% CI, 1.655 to 10.733; P=0.003). Family's consent to donate was significantly associated with head trauma (OR, 3.538; 95% CI, 1.104 to 11.334; P=0.033) and serum sodium level ≥150 mEq (OR, 3.392; 95% CI, 1.404 to 8.194; P=0.007). CONCLUSIONS: Successful organ donation was associated with patient age, systolic blood pressure and serum sodium level. Family's consent to donate was associated with head trauma and serum sodium level.

The Association between Muscle Mass Deficits Estimated from Bioelectrical Impedance Analysis and Lumbar Spine Bone Mineral Density in Korean Adults

BACKGROUND: Bone mineral density (BMD) is influenced by many factors. Despite the reported association between body components and BMD, most of these studies investigated the relationship between absolute muscle mass or fat mass and BMD in postmenopausal women or elderly subjects. The aim of this study is to investigate the association between muscle mass deficits (MMD) estimated from bioelectrical impedance analysis (BIA) and lumbar spinal BMD in Korean adults 20 to 49 years of age. METHODS: This cross-sectional study included 1,765 men and women who visited a health promotion center for a routine checkup. The lumbar spinal BMD was measured by dual energy X-ray absorptiometry. Body composition analysis was performed using BIA. RESULTS: The mean age of the subjects was 40.2±6.3 years. Ten thousand subjects (56.7%) were males and 126 subjects (7.1%) belonged to the low BMD (Z-score ≤-2.0). MMD had the strongest influence on BMD after adjusting for all covariates. The adjusted odds ratio of Group 3 (MDD >2.6 kg) for low BMD was 2.74 (95% CI, 1.46-5.15) after adjusting for age, gender, body mass index, height, and smoking. CONCLUSIONS: MMD estimated by BIA showed a significant association with BMD and could be regarded as an independent risk factor for low BMD in adults 20 to 49 years of age. These findings support that interventions such as physical activity or lifestyle changes may simultaneously modify both muscle and bone health in this age group.

Induction of Autophagy by Low Dose of Cisplatin in H460 Lung Cancer Cells / 결핵및호흡기질환

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.

Induction of Autophagy by Low Dose of Cisplatin in H460 Lung Cancer Cells / 결핵및호흡기질환

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.

The Effect of Autophagy to Cell Death in Nutrient-Deprived H460 Cells / 결핵및호흡기질환

BACKGROUND: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. METHODS: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. RESULTS: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. CONCLUSION: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.

The Effect of Inhibition of Heme Oxygenase-1 on Chemosensitivity of Cisplatin in Lung Cancer Cells / 결핵및호흡기질환

BACKGROUND: Heme oxygenase-1 (HO-1) is known to modulates the cellular functions, including cell proliferation and apoptosis. It is known that a high level of HO-1 expression is found in many tumors, and HO-1 plays an important role in rapid tumor growth on account of its antioxidant and antiapoptotic effects. Cisplatin is a widely used anti-cancer agent for the treatment of lung cancer. However, the development of resistance to cisplatin is a major obstacle to its use in clinical treatment. We previously demonstrated that inhibiting HO-1 expression through the transcriptional activation of Nrf2 induces apoptosis in A549 cells. The aim of this study was to determine of the inhibiting HO-1 enhance the chemosensitivity of A549 cells to cisplatin. MATERIALS AND METHODS: The human lung cancer cell line, A549, was treated cisplatin, and the cell viability was measured by a MTT assay. The change in HO-1, Nrf2, and MAPK expression after the cisplatin treatment was examined by Western blotting. HO-1 inhibition was suppressed by ZnPP, which is a specific pharmacologic inhibitor of HO activity, and small interfering RNA (siRNA). Flow cytometry analysis and Western blot were performed in to determine the level of apoptosis. The level of hydrogen peroxide (H2O2) generation was monitored fluoimetrically using 2',7'-dichlorofluorescein diacetate. RESULTS: The A549 cells showed more resistance to the cisplatin treatment than the other cell lines examined, whereas cisplatin increased the expression of HO-1 and Nrf2, as well as the phosphorylation of MAPK in a time-dependent fashion. Inhibitors of the MAPK pathway blocked the induction of HO-1 and Nrf2 by the cisplatin treatment in A549 cells. In addition, the cisplatin-treated A549 cells transfected with dither the HO-1 small interfering RNA (siRNA) or ZnPP, specific HO-1 inhibitor, showed in a more significantly decrease in viability than the cisplatin-only-treated group. The combination treatment of ZnPP and cisplatin caused in a marked increase in the ROS generation and a decrease in the HO-1 expression. CONCLUSION: Cisplatin increases the expression of HO-1, probably through the MAPK-Nrf2 pathway, and the inhibition of HO-1 enhances the chemosensitivity of A549 cells to cisplatin.