Argonaute 2 (AGO2), the core component of microRNA (miRNA)-induced silencing complex, plays a compelling role in tumorigenesis and aggressiveness. However, the mechanisms regulating the functions of AGO2 in cancer still remain elusive. Herein, we indentify one intronic circular RNA (circRNA) generated from AGO2 gene (circAGO2) as a novel regulator of AGO2-miRNA complexes and cancer progression. CircAGO2 is up-regulated in gastric cancer, colon cancer, prostate cancer, and neuroblastoma, and is associated with poor prognosis of patients. CircAGO2 promotes the growth, invasion, and metastasis of cancer cells in vitro and in vivo. Mechanistic studies reveal that circAGO2 physically interacts with human antigen R (HuR) protein to facilitate its activation and enrichment on the 3'-untranslated region of target genes, resulting in reduction of AGO2 binding and repression of AGO2/miRNA-mediated gene silencing associated with cancer progression. Pre-clinically, administration of lentivirus-mediated short hairpin RNA targeting circAGO2 inhibits the expression of downstream target genes, and suppresses the tumorigenesis and aggressiveness of xenografts in nude mice. In addition, blocking the interaction between circAGO2 and HuR by cell-penetrating inhibitory peptide represses the tumorigenesis and aggressiveness of cancer cells. Taken together, these results indicate that oncogenic circAGO2 drives cancer progression through facilitating HuR-repressed functions of AGO2-miRNA complexes.

BackgroundCircular RNAs (circRNAs), a subclass of non-coding RNAs, play essential roles in tumorigenesis and aggressiveness. Our previous study has identified that circAGO2 drives gastric cancer progression through activating human antigen R (HuR), a protein stabilizing AU-rich element-containing mRNAs. However, the functions and underlying mechanisms of circRNAs derived from HuR in gastric cancer progression remain elusive.MethodsCircRNAs derived from HuR were detected by real-time quantitative RT-PCR and validated by Sanger sequencing. Biotin-labeled RNA pull-down, mass spectrometry, RNA immunoprecipitation, RNA electrophoretic mobility shift, and in vitro binding assays were applied to identify proteins interacting with circRNA. Gene expression regulation was observed by chromatin immunoprecipitation, dual-luciferase assay, real-time quantitative RT-PCR, and western blot assays. Gain- and loss-of-function studies were performed to observe the impacts of circRNA and its protein partner on the growth, invasion, and metastasis of gastric cancer cells in vitro and in vivo.ResultsCirc-HuR (hsa_circ_0049027) was predominantly detected in the nucleus, and was down-regulated in gastric cancer tissues and cell lines. Ectopic expression of circ-HuR suppressed the growth, invasion, and metastasis of gastric cancer cells in vitro and in vivo. Mechanistically, circ-HuR interacted with CCHC-type zinc finger nucleic acid binding protein (CNBP), and subsequently restrained its binding to HuR promoter, resulting in down-regulation of HuR and repression of tumor progression.ConclusionsCirc-HuR serves as a tumor suppressor to inhibit CNBP-facilitated HuR expression and gastric cancer progression, indicating a potential therapeutic target for gastric cancer.

Circular RNAs (circRNA), a subclass of noncoding RNA characterized by covalently closed continuous loops, play emerging roles in tumorigenesis and aggressiveness. However, the functions and underlying mechanisms of circRNA in regulating Wnt/b-catenin signaling and cancer progression remain elusive. Here, we screen cis-acting circRNA generated by b-catenin (CTNNB1)/transcription factor 7like 2 genes and identify one intronic circRNA derived from CTNNB1 (circ-CTNNB1) as a novel driver of cancer progression. Circ-CTNNB1 was predominantly expressed in the nucleus, upregulated in cancer tissues and cell lines, and associated with unfavorable outcomes in patients with cancer. Circ-CTNNB1 promoted b-catenin activation, growth, invasion, and metastasis in cancer cells. Circ-CTNNB1 bound DEAD-box polypeptide 3 (DDX3) to facilitate its physical interaction with transcription factor Yin Yang 1 (YY1), resulting in the transactivation of YY1 and transcriptional alteration of downstream genes associated with b-catenin activation and cancer progression. Preclinically, administration of lentivirus-mediated short hairpin RNA targeting circ-CTNNB1 or a cell-penetrating inhibitory peptide blocking the circ-CTNNB1-DDX3 interaction inhibited downstream gene expression, tumorigenesis, and aggressiveness in cancer cells. Taken together, these results demonstrate cis-acting circ-CTNNB1 as a mediator of b-catenin signaling and cancer progression through DDX3-mediated transactivation of YY1. Significance: These findings reveal the oncogenic functions of a cis-acting circular RNA in b-catenin activation and cancer progression, with potential value as a therapeutic target for human cancers.

Aerobic glycolysis is a hallmark of metabolic reprogramming in tumor progression. However, the mechanisms regulating glycolytic gene expression remain elusive in neuroblastoma (NB), the most common extracranial malignancy in childhood. Herein, we identify that CUT‐like homeobox 1 (CUX1) and CUX1‐generated circular RNA (circ‐CUX1) contribute to aerobic glycolysis and NB progression. Mechanistically, p110 CUX1, a transcription factor generated by proteolytic processing of p200 CUX1, promotes the expression of enolase 1, glucose‐6‐phosphate isomerase, and phosphoglycerate kinase 1, while circ‐CUX1 binds to EWS RNA‐binding protein 1 (EWSR1) to facilitate its interaction with MYC‐associated zinc finger protein (MAZ), resulting in transactivation of MAZ and transcriptional alteration of CUX1 and other genes associated with tumor progression. Administration of an inhibitory peptide blocking circ‐CUX1‐EWSR1 interaction or lentivirus mediating circ‐CUX1 knockdown suppresses aerobic glycolysis, growth, and aggressiveness of NB cells. In clinical NB cases, CUX1 is an independent prognostic factor for unfavorable outcome, and patients with high circ‐CUX1 expression have lower survival probability. These results indicate circ‐CUX1/EWSR1/MAZ axis as a therapeutic target for aerobic glycolysis and NB progression.

Background: Aerobic glycolysis is a hallmark of metabolic reprogramming that contributes to tumor progression. However, the mechanisms regulating expression of glycolytic genes in neuroblastoma (NB), the most common extracranial solid tumor in childhood, still remain elusive. Methods: Crucial transcriptional regulators and their downstream glycolytic genes were identified by integrative analysis of a publicly available expression profiling dataset. In vitro and in vivo assays were undertaken to explore the biological effects and underlying mechanisms of transcriptional regulators in NB cells. Survival analysis was performed by using Kaplan-Meier method and log-rank test. Results: Hepatocyte nuclear factor 4 alpha (HNF4A) and its derived long noncoding RNA (HNF4A-AS1) promoted aerobic glycolysis and NB progression. Gain-and loss-of-function studies indicated that HNF4A and HNF4A-AS1 facilitated the glycolysis process, glucose uptake, lactate production, and ATP levels of NB cells. Mechanistically, transcription factor HNF4A increased the expression of hexokinase 2 (HK2) and solute carrier family 2 member 1 (SLC2A1), while HNF4A-AS1 bound to heterogeneous nuclear ribonucleoprotein U (hnRNPU) to facilitate its interaction with CCCTC-binding factor (CTCF), resulting in transactivation of CTCF and transcriptional alteration of HNF4A and other genes associated with tumor progression. Administration of a small peptide blocking HNF4A-AS1-hnRNPU interaction or lentivirus-mediated short hairpin RNA targeting HNF4A-AS1 significantly suppressed aerobic glycolysis, tumorigenesis, and aggressiveness of NB cells. In clinical NB cases, high expression of HNF4A-AS1, hnRNPU, CTCF, or HNF4A was associated with poor survival of patients. Conclusions: These findings suggest that therapeutic targeting of HNF4A-AS1/hnRNPU/CTCF axis inhibits aerobic glycolysis and NB progression.

The nitrogen (N) budget calculation approach is a useful means of evaluating the impact of human activity on the N cycle. Field scale N budget calculations may ignore the interactions between landscapes, and regional scale calculations rely on statistical data and indirect parameters. Watershed scale budget calculations allow for a more direct quantification of N inputs and outputs. We conducted N budget calculations for a rice paddydominated agricultural watershed in eastern China for 2007-2009, based on intensive monitoring of stream N dynamics, atmospheric deposition, ammonia (NH 3 ) volatilization and household interviews about Nrelated agricultural activities. The results showed that although total N input to the watershed was up to 280 kg N ha -1 year -1 , riverine discharge was only 4.2 kg N ha -1 year -1 , accounting for 1.5% of the total N input, and was further reduced to 2.0 kg N ha -1 year -1 after reservoir storage and/or denitrification removal. The low riverine N output was because of the characteristics of the rice paddydominated landscape, which intercepts run-off and enhances soil denitrification. The watershed actually purified the N in rainwater, as N concentrations in river discharge were much lower than those in rain water. Major N outputs included food/feed export, NH 3 volatilization from chemical fertilizer and manure, and emissions from crop residue burning. Net reactive gaseous emissions (emissions minus deposition) accounted for 5.5% of the total N input, much higher than riverine discharge. Therefore, the agricultural N cycle in such paddy-dominated watersheds impacts the environment mainly through gas exchange rather than water discharge.Keywords Eastern China Á Nitrogen budget Á Reservoir Á Riverine discharge Á Watershed Abbreviations NANI Net anthropogenic nitrogen input N Nitrogen TN Total dissolved nitrogen

De novo design and synthesis of sequence-defined peptoids via amino acid building blocks is reported.

Abstract:This study aimed to statistically and hydrologically assess the performance of the four latest and widely used satellite-gauge combined precipitation estimates (SGPEs), namely CRT (CMORPH CRT), BLD (CMORPH BLD), CDR (PERSIANN CDR), 3B42 (TMPA 3B42 version 7) over the upper yellow river basins (UYRB) in china during 2001-2012 time period. The performances of the SGPEs were compared with the Chinese Meteorological Administration (CMA) datasets using the hydrologic model called Variable Infiltration Capacity (VIC) which is known as a land surface hydrologic model. Results indicated that irrespective of the slight underestimation in the western mountains and overestimation in the southeast, the four SGPEs could generally captured the spatial distribution of precipitation well. Although 3B42 exhibited a better performance in capturing the spatial distribution of daily average precipitation, BLD agreed best with CMA in the time series of watershed average precipitation, which resulted in BLD having a comparable performance to the CMA in the long-term hydrological simulations. Moreover, the potential for disastrous heavy rain mainly occurs in southeastern corner of the basin, and CRT and BLD comparisons showed to be closer to the CMA in the distribution of extreme precipitation events while 3B42 and CDR overestimated the extreme precipitation especially over the southeast of UYRB region. Therefore, CRT and BLD were able to match the high peak discharges very well for the wet seasons, while 3B42 and CDR overrated the high peak discharges. In addition, the four SGPEs performed well for the 2005 flood event but exhibited poorly when tested for the 2012 flood event. Results indicate that the application of the four SGPEs should be used with caution in simulating massive flood events over UYRB region.