Genome-wide expression patterns associated with oncogenesis and sarcomatous transdifferentation of cholangiocarcinoma

BACKGROUND: The molecular mechanisms of CC (cholangiocarcinoma) oncogenesis and progression are poorly understood. This study aimed to determine the genome-wide expression of genes related to CC oncogenesis and sarcomatous transdifferentiation. METHODS: Genes that were differentially expressed between CC cell lines or tissues and cultured normal biliary epithelial (NBE) cells were identified using DNA microarray technology. Expressions were validated in human CC tissues and cells. RESULTS: Using unsupervised hierarchical clustering analysis of the cell line and tissue samples, we identified a set of 342 commonly regulated (>2-fold change) genes. Of these, 53, including tumor-related genes, were upregulated, and 289, including tumor suppressor genes, were downregulated (<0.5 fold change). Expression of SPP1, EFNB2, E2F2, IRX3, PTTG1, PPARγ, KRT17, UCHL1, IGFBP7 and SPARC proteins was immunohistochemically verified in human and hamster CC tissues. Additional unsupervised hierarchical clustering analysis of sarcomatoid CC cells compared to three adenocarcinomatous CC cell lines revealed 292 differentially upregulated genes (>4-fold change), and 267 differentially downregulated genes (<0.25 fold change). The expression of 12 proteins was validated in the CC cell lines by immunoblot analysis and immunohistochemical staining. Of the proteins analyzed, we found upregulation of the expression of the epithelial-mesenchymal transition (EMT)-related proteins VIM and TWIST1, and restoration of the methylation-silenced proteins LDHB, BNIP3, UCHL1, and NPTX2 during sarcomatoid transdifferentiation of CC. CONCLUSION: The deregulation of oncogenes, tumor suppressor genes, and methylation-related genes may be useful in identifying molecular targets for CC diagnosis and prognosis

Microbial Diversity and Volatile Flavor Changes during Gayangju Fermentation, a Traditional Korean House Rice Wine

Physicochemical changes in fermented alcoholic beverages are significantly related to microbial community development during fermentation. Due to its unusually long fermentation, Gayangju, a traditional Korean house rice wine fermented with nuruk as the traditional starter, gives rise to a strong yeast community and, therefore, a high ethanol concentration and different flavors. However, no detailed analysis has been examined. Changes in microbial community structure during Gayangju fermentation were examined using both culture-dependent and culture-independent methods. During fermentation, Saccharomyces cerevisiae and Saccharomycopsis fibuligera were dominant during all stages of the fermentation. In contrast, Candida parapsilosis, Hanseniaspora guilliermondii, Pichia anomala, Malassezia cuniculi and P. fermentans were identified as minor. P. anomala appeared after the second brewing and then remained constant. Among the 19 compounds identified in this study as order-active compounds, 2-methyl-1-butanol (isoamyl alcohol) was the major compound that increased during the long fermentation stage. Most of the odor-active compounds such as 2,3-butanediol, 3-methyl-1-butanol, ethyl tetradecanoate, ethyl decanoate, ethyl dodecanoate, butanoic acid, 3-methylbutanoic acid (isovaleric acid), 2-methylbutanoic acid, 2-methyl-1-propanol, ethyl acetate, ethyl caprylate, 2-phenylethanol, and 3-methylbutyl acetate increased as the fermentation progressed during 68 days of fermentation, which showed significant differences in the concentrations of odor-active compounds of commercially fermented makgeolli

Neuregulin-1 inhibits CoCl2-induced upregulation of excitatory amino acid carrier 1 expression and oxidative stress in SH-SY5Y cells and the hippocampus of mice

Excitatory amino acid carrier 1 (EAAC1) is an important subtype of excitatory amino acid transporters (EAATs) and is the route for neuronal cysteine uptake. CoCl2 is not only a hypoxia-mimetic reagent but also an oxidative stress inducer. Here, we found that CoCl2 induced significant EAAC1 overexpression in SH-SY5Y cells and the hippocampus of mice. Transient transfection of EAAC1 reduced CoCl2-induced cytotoxicity in SH-SY5Y cells. Based on this result, upregulation of EAAC1 expression by CoCl2 is thought to represent a compensatory response against oxidative stress in an acute hypoxic state. We further demonstrated that pretreatment with Neuregulin-1 (NRG1) rescued CoCl2-induced upregulation of EAAC1 and tau expression. NRG1 plays a protective role in the CoCl2-induced accumulation of reactive oxygen species (ROS) and reduction in antioxidative enzyme (SOD and GPx) activity. Moreover, NRG1 attenuated CoCl2-induced apoptosis and cell death. NRG1 inhibited the CoCl2-induced release of cleaved caspase-3 and reduction in Bcl-XL levels. Our novel finding suggests that NRG1 may play a protective role in hypoxia through the inhibition of oxidative stress and thereby maintain normal EAAC1 expression levels.This work was supported by the National Research Foundation of Korea (NRF) and funded by the Ministry of Education, Science and Technology (NRF2019R1H1A2079060 awarded to RSW, NRF-2020R1A2C1011839 awarded to HSK and 2017R1D1A1B03028729 awarded to JHL)