Energy transfer from adipocytes to cancer cells in breast cancer.

Limitations of the current therapeutic approach have raised the need for a novel therapeutic agent in breast cancer. Recently, interest in drugs targeting the tumor microenvironment (TME) had drawn attention in the treatment of breast cancer. Furthermore, recent studies have suggested the role of adipocytes, which are part of the TME, in tumor initiation, growth, and metastasis. In this study, we investigated the metabolic interaction between adipocytes and breast cancer cells and its potential as a new therapeutic target in breast cancer. Breast cancer cell lines and human breast cancer tissue samples were evaluated. Compared to cancer cells cultured alone, or the control group, those co-cultured with adipocytes showed lipid transfer from adipocytes to cancer cells and it was different according to the molecular subtype of breast cancer. Breast cancer cells affected the lipolysis of adipocytes and adipocytes affected the ß-oxidation of breast cancer cells. The key molecule of the process was fatty acid binding protein 4 (FABP4), which is combined with free fatty acid (FFA) and supports its migration to cancer cells. When FABP4 was suppressed, lipid transfer between adipocytes and cancer cells, lipolysis of adipocytes, and ß-oxidation of breast cancer cells were reduced. Furthermore, the expression of lipid metabolism-related proteins and lipolysis-related proteins in breast cancer with adipose stroma showed significantly different expression according to the region of breast cancer tissue. Taken together, we demonstrated the metabolic interaction between adipocytes and breast cancer cells. Breast cancer cells increase the lipolysis in adipocytes and produce a fatty acid, and fatty acid enters into cancer cells. Also, adipocytes contribute to the survival and growth of cancer cells through increased mitochondrial ß-oxidation by using fatty acid from adipocytes. The key molecule of the process is FABP4 and when FABP4 is suppressed, the metabolic interaction is reduced, suggesting its role as a potential therapeutic target.

Expression of DNA methylation-related proteins in metastatic breast cancer.

We aimed to investigate the expression of methylation-related proteins (5-meC and DNMT1) in the metastatic breast cancers of variable sites and its association with clinicopathologic factors. A total of 126 metastatic breast cancers (31 bone metastases, 36 brain metastases, 11 liver metastases, 48 lung metastases) were made into tissue microarray and immunohistochemical staining of ER, PR, HER-2, Ki-67, 5-meC, and DNMT1 were performed. Molecular classification was made on the basis of immunohistochemical staining result of ER, PR, HER-2, Ki-67; luminal A, luminal B, HER-2, triple negative breast cancer (TNBC). Methylation-related proteins were differentially expressed based on the metastatic sites. Tumoral and stromal 5-meC showed the lowest expression in the bone metastasis (P < 0.001), tumoral DNMT1 showed the least expression in bone metastasis and the highest expression in the brain metastasis (P < 0.001). Expression of DNMT1 was correlated with ER negativity (P = 0.004), PR negativity (P = 0.011), HER-2 positivity (P = 0.016), higher Ki-67 labeling indices (P = 0.016), and non-luminal A type (P = 0.017). DNMT1 positivity was associated with shorter overall survival in bone metastasis (P = 0.017) and lung metastasis (P = 0.028) by univariate analysis. In conclusion, methylation-related proteins differentially expressed according to the metastatic sites in metastatic breast cancer. Tumoral and stromal 5-meC showed the lowest expression in the bone metastasis. Tumoral DNMT1 expression was low in bone metastasis and highest in brain metastasis.

Expression of lipid metabolism-related proteins in breast phyllodes tumors.

The aim of this study was to investigate the expression of lipid metabolism-related proteins and the implications thereof in phyllodes tumor (PT) of the breast. A tissue microarray (TMA) was constructed using paraffin blocks from 194 PT patient tissue samples. Immunohistochemical staining for lipid metabolism-related proteins, namely hormone-sensitive lipase (HSL), perilipin 2, fatty-acid-binding proteins 4 (FABP4), carnitine palmitoyltransferase-1 (CPT-1), acyl-CoA oxidase 1 (ACOX-1), and fatty acid synthase (FASN) was performed, and the immunohistochemical staining results were analyzed with respect to clinicopathologic parameters. The numbers of benign, borderline, and malignant PTs were 151, 27, and 16, respectively. The expression of HSL, perilipin 2, FABP4, CPT-1, and FASN in stromal components was higher in higher grade tumors. On univariate analysis, shorter disease-free survival (DFS) was associated with stromal perilipin 2 positivity (p<0.001) and stromal CPT-1 positivity (p=0.004). Shorter overall survival (OS) was associated with stromal perilipin 2 positivity (p<0.001), stromal FABP4 positivity (p<0.001), stromal CPT-1 positivity (p=0.004), and stromal FASN positivity (p<0.001). Multivariate Cox analysis revealed that stromal perilipin 2 positivity (hazard ratio=31.693, 95% CI: 1.341-748.8, p=0.032) was an independent factor for shorter DFS. In conclusion, higher expressions of HSL, perilipin 2, FABP4, CPT-1 and FASN in the stromal component were observed in higher grade PT.

Expression of serine and glycine-related enzymes in phyllodes tumor.

UNLABELLED: Expression patterns of proteins involved in serine and glycine metabolism, and correlations of these patterns with clinicopathologic factors in phyllodes tumor were investigated. Tissue microarrays were prepared from 203 phyllodes tumors (PT) and stained with antibodies specific for glycine decarboxylase (GLDC), phosphoserine aminotransferase 1 (PSAT1), phosphoserine phosphatase (PSPH), phosphoglycerate dehydrogenase (PHGDH), and serine hydroxymethyltransferase 1 (SHMT1). These immunohistochemical results and clinicopathologic parameters were analyzed for correlation. Numbers of benign, borderline, and malignant tumors were 155, 32, and 16, respectively. Stromal expression of PHGDH, PSAT1, PSPH, SHMT1, and GLDC increased with increasing tumor grade, and epithelial expression of SHMT1 also increased with increasing tumor grade (p<0.001, and p=0.005, respectively). On univariate analysis, positive stainings for stromal PHGDH (p<0.001), stromal PSAT1 (p<0.001), stromal PSPH (p=0.003), epithelial SHMT1 (p=0.001), stromal SHMT1 (p=0.022), and stromal GLDC (p<0.001) were each associated with shorter disease-free survival. Stromal GLDC was associated with shorter overall survival (p<0.001). In conclusion, expression of proteins related to serine and glycine metabolism increased with increasing histologic grade in stromal components of phyllodes tumor. KEYWORDS: glycine, tumor grade, metabolism, phyllodes tumor, serine.

Vertebrobasilar dolichoectasia as a risk factor for cerebral microbleeds.

BACKGROUND: Both vertebrobasilar dolichoectasia (VBD) and cerebral microbleeds (CMBs) are related with the risk of intracerebral hemorrhage. We aimed to examine the relationship between the VBD and CMB in ischaemic stroke patients. METHODS: A consecutive series of 182 patients hospitalized because of ischaemic stroke or transient ischaemic attack (TIA), and who underwent gradient echo brain magnetic resonance imaging were retrospectively recruited from a prospective stroke registry. CMB locations were categorized into anterior and posterior circulation. Ectasia was defined as basilar artery (BA) diameter > 4.5 mm, and dolichosis, as either BA bifurcation above the suprasellar cistern or lateral to the margin of the clivus or dorsum sellae. Whether VBD is associated with CMB anywhere in the brain or in anterior or posterior circulation territories was analysed using binary and multinomial logistic regression models. RESULTS: Twenty-four subjects (13.2%) had VBD and 48 (26.4%) had CMBs. CMBs were more frequently observed in patients with VBD than without (66.7% vs. 20.3%, P < 0.001). VBD was significantly associated with CMBs in any location (crude odds ratio, 7.88; 95% confidence interval, 3.10-20.02), in the posterior circulation territory only (9.63; 2.60-34.94), and in both territories (9.25; 3.40-26.29), but not in the anterior circulation only (1.14; 0.009-11.20). These associations remained unchanged after adjusting for age, gender, hypertension, leukoaraiosis and stroke subtype. CONCLUSIONS: VBD in patients with ischaemic stroke or TIA is independently associated with CMBs, especially in the posterior circulation territory.

Comparison of 90-day case-fatality after ischemic stroke between two different stroke outcome registries using propensity score matching analysis.

BACKGROUND: It has not been clarified whether the disparity in ischemic stroke outcome between populations is caused by ethnic and geographic differences or by variations in case mix. Propensity score matching (PSM) analysis can overcome some analytical problems but is rarely used in stroke outcome research. This study was to compare the ischemic stroke case-fatality between two PSM cohorts of Sweden and Korea. METHODS: Prognostic variables related to baseline characteristics and stroke care were included in our PSM model. Then, we selected 7675 Swedish and 1220 Korean patients with ischemic stroke from each stroke registers and performed one-to-one matching based on propensity scores of each patient. RESULTS: After PSM, all measured variables were well balanced in 1163 matched subjects, and the 90-day case-fatality was identical 6.2% (HR 0.997, 95%CI 0.905-1.099) in Sweden and Korea. CONCLUSIONS: No difference is found in the 90-day case-fatality in propensity score-matched Swedish and Korean patients with ischemic stroke.

Androgen receptor expression is significantly associated with better outcomes in estrogen receptor-positive breast cancers.

BACKGROUND: The objective of the study was to evaluate the implications of androgen receptor (AR) in breast cancers. PATIENTS AND METHODS: We investigated immunohistochemical AR expression from the tissue microarrays of 931 patients between 1999 and 2005, and analyzed demographics and outcomes using uni-/multivariate analyses. Tumors with ≥10% nuclear-stained cells were considered positive for AR. RESULTS: AR was expressed in 58.1% of patients. AR was significantly related to older age at diagnosis, smaller size, well-differentiated tumors, higher positivity of hormone receptors, non-triple-negative breast cancers (non-TNBCs), and lower proliferative index. In estrogen receptor (ER)-negative tumors, AR was distinctively associated with human epidermal growth factor receptor type 2 (HER2) overexpression. With a mean follow-up of 72.7 months, AR was positively related to survival in ER-positive but not in ER-negative tumors. In Cox's models, AR was an independent prognostic factor for disease-free survival in ER-positive cancers. Interestingly, molecular apocrine tumors (ER negative and AR positive) with HER2 positive status showed trends of poorer outcome, but AR had no impact on survival in patients with TNBC. CONCLUSIONS: AR is significantly associated with favorable features in breast cancers and related to better outcomes in ER-positive not in ER-negative tumors. These results suggest that AR could be an additional marker for endocrine responsiveness in ER-positive cancers and a candidate for therapeutic targeting of ER-negative tumors.

EGFR and HER-2 status of non-small cell lung cancer brain metastasis and corresponding primary tumor.

We investigated EGFR and HER-2 status in brain metastatic non-small cell lung cancer (NSCLC) and compared them to EGFR and HER-2 status of primary NSCLC. Evaluated were 66 cases of brain metastatic NSCLC, including 20 cases of corresponding primary NSCLC. HER-2 status was investigated by immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH), and EGFR status was evaluated by IHC. HER-2 overexpression and/or amplification was/were observed in three cases (4.5 %) of 66 cases of brain metastatic NSCLC, and 23 cases (34.8%) demonstrated EGFR overexpression. Among 20 cases of primary and corresponding metastatic NSCLC, one case showed HER-2 overexpression and amplification in both primary and metastatic tumor. On the other hand, EGFR overexpression was noted in four cases of primary NSCLC and nine cases of metastatic NSCLC. Five cases showed EGFR gain in metastatic NSCLC. Brain metastatic NSCLC demonstrated different expression patterns of the abovementioned biomarkers, particularly EGFR when compared to primary NSCLC. Therefore, HER-2 and EGFR status are suggested to be evaluated in brain metastatic NSCLC for targeted monotherapy.

Effect of cilnidipine vs losartan on cerebral blood flow in hypertensive patients with a history of ischemic stroke: a randomized controlled trial.

OBJECTIVES: The aim of this study was to compare the effects of antihypertensive agents on cerebral blood flow (CBF) in hypertensive patients with previous ischemic stroke. MATERIALS AND METHODS: In this double-blind, multi-center, non-inferiority trial, 196 patients were randomized to cilnidipine 10-20 mg or losartan 50-100 mg once daily for 4 weeks. Baseline and follow-up CBF as measured by single photon emission computed tomography were obtained in 167. The primary endpoint was the global CBF change. The secondary endpoints were the CBF change in the hemisphere ipsilateral to the index stroke, non-impairment of global CBF and blood pressure (BP) reduction. RESULTS: Global CBF increased significantly in the cilnidipine arm (9.0 +/- 29.6%, P = 0.0071) and the losartan arm (11.4 +/- 31.4%, P = 0.0012), and these changes were not different between the two groups (P = 0.607). However, the estimated difference in percentage global CBF change between the two groups was -2.43% (97.5% CI, -13.06% to 8.21%), which crossed the predetermined non-inferiority margin of -8.6%. Ipsilesional hemispheric CBF change, non-impairment of global CBF and BP reduction were similar in the two groups. CONCLUSIONS: This trial failed to prove the non-inferiority of cilnidipine to losartan regarding global CBF change. Both the treatments, however, increase the global CBF despite BP lowering.

Efficacy of Helicobacter pylori eradication therapy in chronic liver disease.

BACKGROUND/AIMS: Peptic ulcers occur more commonly in patients with liver cirrhosis (LC). Helicobacter pylori is recognized as the most important etiology in the pathogenesis of peptic ulcers. We investigated the efficacy of proton pump inhibitor (PPI)-based triple therapy in patients with chronic liver disease and peptic ulcer. PATIENTS AND METHODS: One hundred sixty-three patients with LC or chronic hepatitis (CH) with a peptic ulcer and proven H. pylori infection were included. The combination of PPI, amoxicillin (1.0 g), and clarithromycin (500 mg), each given twice daily, was administered for 1 or 2 weeks. The eradication of H. pylori was determined by the rapid urease test, histology, or the 13C-urea breath test at least 4 weeks after completing the treatment. RESULTS: The eradication rate of H. pylori was similar between the LC and CH groups; 82.6% and 88.1%, respectively. In addition, there were no significant differences in eradication rates between the patients with Child-Pugh class A and Child-Pugh class B/C disease. The side effects in each group were generally mild. Only the serum ALT levels showed a significant correlation with the success of H. pylori eradication in both the LC and CH groups. CONCLUSION: The PPI-based triple therapy achieves high eradication rates for H. pylori infection, in patients with chronic liver disease, without significant side effects.

Impact of neurological and medical complications on 3-month outcomes in acute ischaemic stroke.

OBJECTIVE: To evaluate the impact of neurological and medical complications on 3-month outcomes in acute ischaemic stroke patients. METHODS: We prospectively investigated complications for all the consecutive acute ischaemic stroke patients admitted within 7 days from onset in four university hospitals during a 1-year period. Baseline data and 3-month outcomes were collected. Poor outcome was defined as a modified Rankin Scale score 3-6. RESULTS: A total of 1 254 patients were recruited: 264 (21.1%) and 303 (24.2%) patients experienced one or more neurological and medical complications, respectively. The most common complications were ischaemic stroke progression (17.1%) and pneumonia (12.0%). Of 1 233 patients with available 3-month outcomes, 34.9% had a poor outcome. Multivariate analysis revealed that neurological (odds ratio, 95% confidence interval; 5.47, 3.63-8.24) and medical (3.47, 2.30-5.23) complications were independent predictors of the poor outcome. For the individual complications, ischaemic stroke progression (7.48, 4.73-11.84), symptomatic hemorrhagic transformation (3.57, 1.33-9.54), pneumonia (4.44, 2.20-8.99), extracranial bleeding (4.45, 1.88-10.53), and urinary tract infection (2.72, 1.32-5.60) were independently associated with the poor outcome. CONCLUSION: Outcome after ischaemic stroke is adversely influenced by complications, especially ischaemic stroke progression, symptomatic hemorrhagic transformation, pneumonia, extracranial bleeding, and urinary tract infection. Interventions to prevent those complications might improve ischaemic stroke outcome.

APOE epsilon2/epsilon4 polymorphism and cerebral microbleeds on gradient-echo MRI.

The association of APOE genotypes with cerebral microbleeds (CMBs) was examined on the basis of the location of CMBs in 414 patients who were admitted primarily because of stroke. With respect to possession of the epsilon2 or epsilon4 allele, the adjusted odds ratio was 1.94 (1.05 to 3.58) for lobar CMBs but 1.21 (0.69 to 2.11) for nonlobar CMBs. This suggests that the pathogenesis of CMBs may differ depending on their location.

Evaluation of bipolar electrocoagulation applied to biofiltration for phosphorus removal.

To reduce the residual organic matter and phosphorus contained in secondary effluent, a biofiltration system combined with electrocoagulation using bipolar iron electrodes was evaluated as a supplementary treatment to existing small-community sewage treatment. Based on the results of batch tests, bipolar electrocoagulation (BEC) was found to be more effective on phosphorus removal than monopolar electrocoagulation (MEC) but energy consumption was less in monopolar electrocoagulation. Optimum conditions of BEC to treat the secondary effluent were current density 15 A/m2, electrode spacing 1 cm and pH < 8. The removals of COD(Cr) and phosphorus by biofiltration system without BEC were 69.1% and 9.6%, respectively. However, biofiltration system combined with BEC showed 76.6-83.7% and 70.7-93.0% removal for COD(Cr) and phosphorus respectively. Extraordinary increase in phosphorus could be achieved by introducing electrocoagulation to biofiltration, and BEC/biofiltration system was evaluated to be applicable to existing small-community sewage treatment plants as a supplementary process.

Global detection of molecular changes reveals concurrent alteration of several biological pathways in nonsmall cell lung cancer cells.

To identify the molecular changes that occur in non-small cell lung carcinoma (NSCLC), we compared the gene expression profile of the NCI-H292 (H292) NSCLC cell line with that of normal human tracheobronchial epithelial (NHTBE) cells. The NHTBE cells were grown in a three-dimensional organotypic culture system that permits maintenance of the normal pseudostratified mucociliary phenotype characteristic of bronchial epithelium in vivo. Microarray analysis using the Affymetrix oligonucleotide chip U95Av2 revealed that 1,683 genes showed a >1.5-fold change in expression in the H292 cell line relative to the NHTBE cells. Specifically, 418 genes were downregulated and 1,265 were upregulated in the H292 cells. The expression data for selected genes were validated in several different NSCLC cell lines using quantitative real-time PCR and Western analysis. Further analysis of the differentially expressed genes indicated that WNT responses, apoptosis, cell cycle regulation and cell proliferation were significantly altered in the H292 cells. Functional analysis using fluorescence-activated cell sorting confirmed concurrent changes in the activity of these pathways in the H292 line. These findings show that (1) NSCLC cells display deregulation of the WNT, apoptosis, proliferation and cell cycle pathways, as has been found in many other types of cancer cells, and (2) that organotypically cultured NHTBE cells can be used as a reference to identify genes and pathways that are differentially expressed in tumor cells derived from bronchogenic epithelium.

Regulation of mucous differentiation and mucin gene expression in the tracheobronchial epithelium.

The goal of our studies is to elucidate mechanisms that control and modulate mucous differentiation and mucin gene expression in the conducting airways. We used cultures of normal human tracheobronchial epithelial (NHTBE) cells that were shown to secrete two major airway mucins, namely MUC5AC and MUC5B as well as several other secretory products. Mucous differentiation and expression of MUC2, MUC5AC, MUC5B and MUC7, but not MUCi, MUC4, and MUC8 mucin genes, were shown to be retinoic acid- (RA) or retinol-dependent. We found that RA control of mucin genes was mediated by the retinoid acid receptors RAR alpha and, to a lesser extent, by RAR gamma. Our studies also showed that other important bioregulators such as thyroid hormone (T3) and epidermal growth factor (EGF) modulate basal expression of mucin genes, interacting with RA in a concentration-dependent manner. T3, which binds to thyroid receptors (TRs) belonging to the same superfamily of steroid hormone nuclear receptors as the RARs, inhibits mucin gene expression, particularly MUC5AC. One possible mechanism of this T3 effect is downregulation of RAR proteins, which are critical for mucin gene expression. However, we also found that T3 inhibits MUC5AC transcription.EGF, which had previously been shown to stimulate mucin expression and mucin secretion in cultured rat tracheal epithelial (RTE) cells, inhibited mucin secretion in human bronchial epithelial cell cultures. This effect was EGF concentration- and time-dependent and was progressively abolished by increasing the RA concentration. Subsequent studies suggested that the inhibitory effects of high concentrations of EGF may result from selective reduction of MUC5AC expression. These studies thus point to potentially important species differences in the mechanisms regulating mucous production, and they also confirm previous findings indicating differential regulation of MUC5AC and MUC5B gene expression.

Vanadium stimulates human bronchial epithelial cells to produce heparin-binding epidermal growth factor-like growth factor: a mitogen for lung fibroblasts.

The bronchial epithelium is a potential source of growth factors that could mediate airway fibrosis during the progression of diseases such as asthma and chronic bronchitis. We report that conditioned medium (CM) from normal human bronchial epithelial cells (NHBECs) contains mitogenic activity for human lung fibroblasts that is blocked by the epidermal growth factor receptor (EGF-R) tyrosine kinase inhibitor AG1478 and by neutralizing antibodies raised against heparin-binding epidermal growth factor-like growth factor (HB-EGF). Neutralizing antibodies against other EGF-R ligands (EGF and transforming growth factor-alpha) or other antibodies against growth factors (platelet-derived growth factors, insulin-like growth factor-1) had no affect on the mitogenic activity of NHBEC-CM. HB-EGF messenger RNA (mRNA) expression in NHBEC was detected by reverse transcriptase/polymerase chain reaction and Northern blot analysis. HB-EGF protein was detected by enzyme-linked immunosorbent assay. Vanadium pentoxide (V2O5), a fibrogenic metal associated with occupational asthma, caused a several-fold increase in HB-EGF mRNA expression and protein, whereas the inert metal titanium dioxide had no effect on HB-EGF expression. V2O5-induced HB-EGF mRNA expression was inhibited by the EGF-R tyrosine kinase inhibitor AG1478, the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580, and the MAP kinase kinase inhibitor PD98059. Finally, HB-EGF induced the production of fibroblast growth factor (FGF)-2 by human lung fibroblasts and anti-FGF-2 antibody partially blocked the mitogenic activity of NHBEC-CM on fibroblasts. These data suggest that HB-EGF is a fibroblast mitogen produced by NHBECs and that induction of an FGF-2 autocrine loop in fibroblasts by HB-EGF accounts for part of this mitogenic activity.