Multifactorial anticancer effects of digalloyl-resveratrol encompass apoptosis, cell-cycle arrest, and inhibition of lymphendothelial gap formation in vitro

BACKGROUND: Digalloyl-resveratrol (di-GA) is a synthetic compound aimed to combine the biological effects of the plant polyhydroxy phenols gallic acid and resveratrol, which are both radical scavengers and cyclooxygenase inhibitors exhibiting anticancer activity. Their broad spectrum of activities may probably be due to adjacent free hydroxyl groups. METHODS: Protein activation and expression were analysed by western blotting, deoxyribonucleoside triphosphate levels by HPLC, ribonucleotide reductase activity by 14 C-cytidine incorporation into nascent DNA and cell-cycle distribution by FACS. Apoptosis was measured by Hoechst 33258/propidium iodide double staining of nuclear chromatin and the formation of gaps into the lymphendothelial barrier in a three-dimensional co-culture model consisting of MCF-7 tumour cell spheroids and human lymphendothelial monolayers. RESULTS: In HL-60 leukaemia cells, di-GA activated caspase 3 and dose-dependently induced apoptosis. It further inhibited cell-cycle progression in the G1 phase by four different mechanisms: rapid downregulation of cyclin D1, induction of Chk2 with simultaneous downregulation of Cdc25A, induction of the Cdk-inhibitor p21(Cip/Waf) and inhibition of ribonucleotide reductase activity resulting in reduced dCTP and dTTP levels. Furthermore, di-GA inhibited the generation of lymphendothelial gaps by cancer cell spheroid-secreted lipoxygenase metabolites. Lymphendothelial gaps, adjacent to tumour bulks, can be considered as gates facilitating metastatic spread. CONCLUSION: These data show that di-GA exhibits three distinct anticancer activities: induction of apoptosis, cell-cycle arrest and disruption of cancer cell-induced lymphendothelial disintegration. British Journal of Cancer (2010) 102, 1361-1370. doi:10.1038/sj.bjc.6605656 www.bjcancer.com (C) 2010 Cancer Research U

Texture-based Automated Quantification of Interstitial Lung Disease: Correlation With the Visual Score

Purpose of the Study: The quantification of various disease patterns in the lung parenchyma remains a challenge. In this study the texture analysis algorithm 3D-AMFM (Adaptive Multiple Feature Method) contained in the software PASS (University of Iowa) was applied with interstitial lung disease (ILD). We checked for the statistical accuracy and reliabity of the method by standard tests compared to visual scoring. Methods: Based on a Bayesian classifier, a training data base including texture patterns (normal, ground glass, honey combing, emphysema) from 1300 volumes of interest (VOIs; 151515 pixels) of 47 selected patients with mixed ILDs was built up. Another 18 patients with a typical thin-section CT pattern of usual interstitial pneumonia (UIP) (n=9) and nonspecific interstitial pneumonia (NSIP) (n=9) were independently analyzed and visually quantified at 5 pre-established levels by two experienced chest radiologists. The same thin-section CT scans were analyzed with 3D-AMFM. Wilcoxon test was used to evaluate the correlation between the visual scores and the computed results. Results: The mean extent of honeycombing, ground glass and emphysema was 5.4%, 43.5% and 2.1% by the visual score and 19.4%, 44.3% and 0.6% by the 3D-AMFM, respectively. There was close correlation between visual score and 3D-AMFM for both the extent of ground glass (P=0.546) and emphysema (P=0.099), but worse for the extent of honey combing (P=0.000837). Conclusions: The 3D-AMFM system is a promising and effective tool for ILD quantification, showing clinical acceptable correlation with human observer. The overestimation of honeycombing by 3D-AMFM is probably caused by small vessels and airways. The continuing development of the feature data base and the inclusion of further pathologic texture patterns will improve quantification of disease and provide objective measures of disease progression

In vitro inhibition of breast cancer spheroid-induced lymphendothelial defects resembling intravasation into the lymphatic vasculature by acetohexamide, isoxsuprine, nifedipin and proadifen.

BACKGROUND: As metastasis is the prime cause of death from malignancies, there is vibrant interest to discover options for the management of the different mechanistic steps of tumour spreading. Some approved pharmaceuticals exhibit activities against diseases they have not been developed for. In order to discover such activities that might attenuate lymph node metastasis, we investigated 225 drugs, which are approved by the US Food and Drug Administration. METHODS: A three-dimensional cell co-culture assay was utilised measuring tumour cell-induced disintegrations of the lymphendothelial wall through which tumour emboli can intravasate as a limiting step in lymph node metastasis of ductal breast cancer. The disintegrated areas in the lymphendothelial cell (LEC) monolayers were induced by 12(S)-HETE, which is secreted by MCF-7 tumour cell spheroids, and are called 'circular chemorepellent induced defects' (CCIDs). The putative mechanisms by which active drugs prevented the formation of entry gates were investigated by western blotting, NF-κB activity assay and by the determination of 12(S)-HETE synthesis. RESULTS: Acetohexamide, nifedipin, isoxsuprine and proadifen dose dependently inhibited the formation of CCIDs in LEC monolayers and inhibited markers of epithelial-to-mesenchymal-transition and migration. The migration of LECs is a prerequisite of CCID formation, and these drugs either repressed paxillin levels or the activities of myosin light chain 2, or myosin-binding subunit of myosin phosphatase. Isoxsuprine inhibited all three migration markers, and isoxsuprine and acetohexamide suppressed the synthesis of 12(S)-HETE, whereas proadifen and nifedipin inhibited NF-κB activation. Both the signalling pathways independently cause CCID formation. CONCLUSION: The targeting of different mechanisms was most likely the reason for synergistic effects of different drug combinations on the inhibition of CCID formation. Furthermore, the treatment with drug combinations allowed also a several-fold reduction in drug concentrations. These results encourage further screening of approved drugs and their in vivo testing