Background:Functional microRNAs (miRNAs) in exosomes have been recognised as potential stable biomarkers in cancers. The aim of this study is to identify specific miRNAs in exosome as serum biomarkers for the early detection of recurrence in human colorectal cancer (CRC).Methods:Serum samples were sequentially obtained from six patients with and without recurrent CRC. The miRNAs were purified from exosomes, and miRNA microarray analysis was performed. The miRNA expression profiles and copy number aberrations were explored using microarray and array CGH analyses in 124 CRC tissues. Then, we validated exosomal miRNAs in 2 serum sample sets (90 and 209 CRC patients) by quantitative real-time RT–PCR.Results:Exosomal miR-17-92a cluster expression level in serum was correlated with the recurrence of CRC. Exosomal miR-19a expression levels in serum were significantly increased in patients with CRC as compared with healthy individuals with gene amplification. The CRC patients with high exosomal miR-19a expression showed poorer prognoses than the low expression group (P<0.001).Conclusions:Abundant expression of exosomal miR-19a in serum was identified as a prognostic biomarker for recurrence in CRC patients.

Background:We previously conducted gene expression microarray analyses to identify novel indicators for colorectal cancer (CRC) metastasis and prognosis from which we identified PVT-1 as a candidate gene. PVT-1, which encodes a long noncoding RNA, mapped to chromosome 8q24 whose copy-number amplification is one of the most frequent events in a wide variety of malignant diseases. However, PVT-1 molecular mechanism of action remains unclear.Methods:We conducted cell proliferation and invasion assays using colorectal cancer cell lines transfected with PVT-1siRNA or negative control siRNA. Gene expression microarray analyses on these cell lines were also carried out to investigate the molecular function of PVT-1. Further, we investigated the impact of PVT-1 expression on the prognosis of 164 colorectal cancer patients by qRT–PCR.Results:CRC cells transfected with PVT-1 siRNA exhibited significant loss of their proliferation and invasion capabilities. In these cells, the TGF-β signalling pathway and apoptotic signals were significantly activated. In addition, univariate and multivariate analysis revealed that PVT-1 expression level was an independent risk factor for overall survival of colorectal cancer patients.Conclusion:PVT-1, which maps to 8q24, generates antiapoptotic activity in CRC, and abnormal expression of PVT-1 was a prognostic indicator for CRC patients.

BackgroundPredictive biomarkers for the recurrence of hepatocellular carcinoma (HCC) have great benefit in the selection of treatment options, including liver transplantation (LT), for HCC. The purpose of this study was to identify specific microRNAs (miRs) in exosomes from the serum of patients with recurrent HCC and to validate these molecules as novel biomarkers for HCC recurrence.MethodsWe employed microarray-based expression profiling of miRs derived from exosomes in the serum of HCC patients to identify a biomarker that distinguishes between patients with and without HCC recurrence after LT. This was followed by the validation in a separate cohort of 59 HCC patients who underwent living related LT. The functions and potential gene targets of the recurrence-specific miRs were analysed using a database, clinical samples and HCC cell lines.ResultsWe found that miR-718 showed significantly different expression in the serum exosomes of HCC cases with recurrence after LT compared with those without recurrence. Decreased expression of miR-718 was associated with HCC tumour aggressiveness in the validated cohort series. We identified HOXB8 as a potential target gene of miR-718, and its upregulation was associated with poor prognosis.ConclusionCirculating miRs in serum exosomes have potential as novel biomarkers for predicting HCC recurrence.

NEK2 may be an independent prognostic factor for CRC and was regulated by miR-128, a microRNA that was subjected to epigenetic regulation. Thus, this miR-128/NEK2 pathway may be a prospective therapeutic target for patients with CRC.

Background:Paired related homoeobox 1 (PRRX1) has been identified as a new epithelial-mesenchymal transition (EMT) inducer in breast cancer. However, the function of PRRX1 in colorectal cancer (CRC) has not been elucidated.Methods:We utilised ectopic PRRX1-expressing cell lines to analyse the function of PRRX1 in CRC. The clinical significance of PRRX1 was also examined on three independent CRC case sets.Results:PRRX1 induced EMT and the stem-like phenotype in CRC cells. In contrast to studies of breast cancer, abundant expression of PRRX1 was significantly associated with metastasis and poor prognosis in CRC.Conclusion:PRRX1 is an indicator of metastasis and poor prognosis in CRC cases. Further investigation is required to uncover the signalling network regulating PRRX1.

The chromodomain helicase DNA-binding (CHD) family comprises a class of chromatin remodeling enzymes. Previous studies suggest that CHD8 may negatively regulate various genes and signaling pathways, such as the Wnt/β‑catenin pathway. However, few studies have investigated the role of CHD8 in cancer cells. We analyzed the expression of CHD8 in cancer lesions and corresponding non-cancerous tissues to demonstrate the prognostic significance of CHD8 expression in 101 cases of gastric cancer. We also investigated the functional implications of aberrant CHD8 expression by conducting gene set enrichment analysis (GSEA). Expression of CHD8 mRNA was significantly lower in gastric cancer tissues compared to that in corresponding normal tissues (P=0.003). In multivariate analysis for overall survival, we found that CHD8 expression was an independent prognostic factor in gastric cancer. Moreover, GSEA revealed that CHD8 was significantly associated with genes involved in the Wnt/β‑catenin pathway and in the cell cycle. In addition, knockdown of CHD8 expression in the gastric cancer cell lines, MKN45 and NUGC4, promoted proliferation. In conclusion, the present study suggests that loss of CHD8 expression may be a novel indicator for biological aggressiveness in gastric cancer.

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1. Infusion of 2 mM ethanol into perfused liver from fed rats increased the rate of oxygen uptake concomitant with the decrease in the rate of glycolysis (lactate + pyruvate production). A linear correlation (r = 0.92) was observed between the increase in the rate of oxygen uptake and the decrease in the rate of lactate + pyruvate production determined on the whole organ by the difference between influent minus effluent concentration.2. Miniature oxygen electrodes (tip diameter, 50 pm) were then placed on periportal or pericentral regions of the lobule on the liver surface, and local rates of oxygen uptake were determined by stopping the flow of perfusate and monitoring the rate of decrease of oxygen concentration ('stopped-flow oxygen uptake technique'). During perfusion in the anterograde direction, ethanol infusion (2 mM) increased rates of oxygen uptake about twofold more in pericentral (15 pmol x g-x h-') than in periportal (7 pmol x g-' x h-') regions of the liver lobule in livers from well-fed rats. In contrast, ethanol did not affect the rate of oxygen uptake significantly in either region of the liver lobule in livers from fasted rats.3. Glucose (30 mM) decreased oxygen concentrations initially in both regions of the liver lobule when infused into livers from fasted rats perfused in the anterograde direction. Subsequently, glucose increased the oxygen concentration in pericentral but not periportal regions of the liver lobule. This increase in regional oxygen concentration correlated temporally (r = 0.99) with increases in rates of glycolysis. The addition of ethanol in the presence of glucose reduced the rate of lactate + pyruvate production and increased the rate of oxygen uptake predominantly in pericentral regions.4. These data are consistent with the following interpretation. Ethanol metabolism elevates NADH in both periportal and pericentral regions of the liver lobule causing redox inhibition of glyceraldehyde-3-phosphate dehydrogenase and decreased rates of glycolytic ATP synthesis. The ADP not phosphorylated in the cytosol then moves into the mitochondrion and stimulates oxygen uptake. Since ethanol and glucose elevated oxygen uptake to a greater extent in pericentral regions of the liver lobule, it is concluded that glycolysis predominates in hepatocytes located proximal to the central vein during perfusion in the anterograde direction.5. When similar experiments were performed with perfusion in the retrograde direction, glycolysis was localized in periportal regions of the liver lobule. Thus, metabolic zonation of carbohydrate metabolism can be shifted rapidly within regions of the liver lobule in perfused liver.For the last two decades evidence has accumulated, mainly from histochemical and microchemical studies, indicating that several key enzymes and metabolites of carbohydrate metabolism are distributed unevenly in different zones of the liver lobule [l -31. Based on these differences and the fact that the zones shifted dynamically during the feeding-fasting cycle, Jungermann and his coll...