Characterization of bitter-tasting and antioxidant activity of dry-hopped beers.

BACKGROUND: Bitter flavors and antioxidant activities are critical characteristics and indicators, respectively, of beer quality. To gain a better understanding of dry-hopped beer's bitterness, this work comprehensively evaluated the perceived bitterness intensity and bitterness attributes from aspects of beer aroma and non-volatile bitter compounds using sensory analysis under two conditions: (i) with and (ii) without nose clips. To quantify bitter and volatile compounds, the work conducted chromatographic analyses: high-performance liquid chromatography (HPLC), ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and gas chromatography-mass spectrometry (GC-MS). Simultaneously, this work assessed the antioxidant activity of commercially dry-hopped beers. RESULTS: First, dry-hopped beer in this study contained abundant non-volatile bitter compounds (hop bitter acids, polyphenols and flavonoids), and aroma was validated using HPLC, UPLC-MS and GC-MS methods. Moreover, the bitter-tasting perception test findings demonstrated that many dry-hopped beers had a higher bitterness intensity when evaluated without a nose clip, whereas this phenomenon was adverse in several ale beers. Additionally, the 'lingering' and 'harsh' characteristics were diminished when beer aroma was blocked out (with nose clip) for dry-hopped beer. Meanwhile, most dry-hopped beers had greater antioxidant activity than ale beers (P < 0.05). CONCLUSION: This work revealed the bitterness complexity of dry-hopped beer; besides non-volatile bitter compounds, beer aroma had an impact on the perceived bitterness intensity and attributes, and dry-hopped beer had a relatively intense antioxidant capacity. This study facilitated the development of a detailed knowledge about the assessment of bitter-tasting perceptions in dry-hopped beers and provided a basis for the development of functional beer benefiting human health. © 2022 Society of Chemical Industry.

TMSB10, a potential prognosis prediction biomarker, promotes the invasion and angiogenesis of gastric cancer.

BACKGROUND AND AIM: The thymosin beta 10 (TMSB10) was originally identified from the thymus, which plays a key role in the development of many cancers. However, the underlying molecular mechanisms of TMSB10 involved in GC have not been understood. METHODS: We sought to determine the expression of TMSB10 in human GC tissues and illustrate whether it is correlated with the clinical pathologic characteristics and prognosis in GC patients. Its roles and potential mechanisms in regulating tumor growth, invasion, and angiogenesis were evaluated by TMSB10 knockdown/overexpression of GC cells in vitro and ex vivo. RESULTS: Marked overexpression of TMSB10 protein expression was observed in GC cells and tissues, which was associated with the advanced tumor stage and lymph nodes (LN) metastasis of GC patients. Furthermore, prognostic analysis showed that GC patients with high TMSB10 expression had a remarkably shorter survival and acted as an important factor for predicting poor overall survival in GC patients. Moreover, TMSB10 overexpression promoted, while TMSB10 knockdown the proliferation, EMT process, and angiogenesis of GC cells. CONCLUSION: The study highlights that TMSB10 may hold promise as potential prognosis prediction biomarker for the diagnosis of GC and a potential therapeutic target, which will facilitate the development of a novel therapeutic strategy against GC.

Contamination, distribution, and risk assessment of antibiotics in the urban surface water of the Pearl River in Guangzhou, South China.

To assess the impact of antibiotic pollution to the ecosystem in urban water, the occurrence, seasonal, and spatial distributions, potential sources, and ecological risks of 18 targeted antibiotics in urban river, Pearl River located in Guangzhou city, were investigated. Surface water samples were sampled from 24 sites in Guangzhou center of Pearl River during dry and wet seasons. The results indicated that the concentrations of antibiotic residues were at the nanogram per liter level, except sulfamethazine (SMD) (µg/L). Sulfonamides (SAs) were the dominant antibiotics, contributing 60.4-65.0% to the total antibiotics. The concentrations of SAs, fluoroquinolones (QUs), macrolides (MLs), tetracyclines (TCs), and lincosamides (LCs) were higher in dry season than those in wet season at most sampling sites, which possibly resulted from the dilution effect of heavy rainfall. The concentrations of the antibiotic residues in Guangzhou were comparable or higher than other urban rivers. The calculation on risk quotients indicated that erythromycin-H2O (ETM-H2O) and tetracycline (TC) were of high risks. The source identification by the Pearson correlation analysis and principal component analysis-multiple linear regression (PCA-MLR) method suggested that municipal wastewater treatment plants were primary sources of antibiotics. These results would provide important information for the environmental protect.

Inhibition of carnitine palmitoyl transferase 1A-induced fatty acid oxidation suppresses cell progression in gastric cancer.

BACKGROUND: Gastric cancer (GC) has a high rate of metastasis which thereason leading to death. Carnitine palmitoyl transferase 1a (CPT1A) has been reported to play a critical obstacle to various types of cancer progression, which is an attractive focus in anti-cancer therapy. However, the underlying molecular mechanisms of CPT1A involved in GC have not been clarified clear. METHODS: To determine the expression of CPT1A in human GC tissues and cells and illustrate whether it is correlated with the clinical pathologic characteristics and prognosis in GC patients. Its roles and potential mechanisms in regulating tumor growth and invasion were evaluated by CPT1A knockdown/overexpression of GC cells in vitro. RESULTS: Marked upregulation of CPT1A protein expression was observed in GC cells and tissues, which was associated with grade, pathological stage, lymph node metastasis and poor prognosis in patients with GC. CPT1A overexpression also promoted the proliferation, invasion, EMT process of GC cells. In addition, CPT1A upregulation activated GC cell fatty acid oxidation (FAO) via increasing NADP+/NADPH ratio, whereas inhibiting of FAO abolished the effects of CPT1A on GC cell proliferation and migration. CONCLUSION: Our results examine that CPT1A-mediated FAO activation increases GC cell proliferation and migration, supporting that CPT1A is a useful prognostic biomarker and an attractive focus for GC.

Targeted gene editing restores regulated CD40L function in X-linked hyper-IgM syndrome.

Loss of CD40 ligand (CD40L) expression or function results in X-linked hyper-immunoglobulin (Ig)M syndrome (X-HIGM), characterized by recurrent infections due to impaired immunoglobulin class-switching and somatic hypermutation. Previous attempts using retroviral gene transfer to correct murine CD40L expression restored immune function; however, treated mice developed lymphoproliferative disease, likely due to viral-promoter-dependent constitutive CD40L expression. These observations highlight the importance of preserving endogenous gene regulation in order to safely correct this disorder. Here, we report efficient, on-target, homology-directed repair (HDR) editing of the CD40LG locus in primary human T cells using a combination of a transcription activator-like effector nuclease-induced double-strand break and a donor template delivered by recombinant adeno-associated virus. HDR-mediated insertion of a coding sequence (green fluorescent protein or CD40L) upstream of the translation start site within exon 1 allowed transgene expression to be regulated by endogenous CD40LG promoter/enhancer elements. Additionally, inclusion of the CD40LG 3'-untranslated region in the transgene preserved posttranscriptional regulation. Expression kinetics of the transgene paralleled that of endogenous CD40L in unedited T cells, both at rest and in response to T-cell stimulation. The use of this method to edit X-HIGM patient T cells restored normal expression of CD40L and CD40-murine IgG Fc fusion protein (CD40-muIg) binding, and rescued IgG class switching of naive B cells in vitro. These results demonstrate the feasibility of engineered nuclease-directed gene repair to restore endogenously regulated CD40L, and the potential for its use in T-cell therapy for X-HIGM syndrome.

Lysinibacillus manganicus sp. nov., isolated from manganese mining soil.

A Gram-stain-positive, aerobic, motile, rod-shaped bacterium, designated strain Mn1-7(T), was isolated from manganese mining soil in Tianjin, China. The closest phylogenetic relatives were Lysinibacillus massiliensis CCUG 49529(T) (97.2 % 16S rRNA gene sequence similarity), L. xylanilyticus XDB9(T) (96.7 %), L. sinduriensis JCM 15800(T) (96.2 %), L. odysseyi NBRC 100172(T) (95.9 %) and L. boronitolerans NBRC 103108(T) (95.4 %) (the type species of the genus). DNA-DNA hybridization values for strain Mn1-7(T) with the type strains of L. massiliensis and L. sinduriensis were 24.9 and 27.7 %, respectively. The genomic DNA G+C content was 38.4 mol%. The major menaquinone was MK-7 and the major fatty acids were iso-C15 : 0, iso-C16 : 0 and iso-C14 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The cell-wall peptidoglycan was type A4α (L-Lys-D-Asp), and the predominant cell-wall sugar was xylose. DNA-DNA hybridization results and comparison of phenotypic and chemotaxonomic characters between strain Mn1-7(T) and the phylogenetically most closely related strains revealed that the isolate represents a novel species of the genus Lysinibacillus, for which the name Lysinibacillus manganicus sp. nov. is proposed. The type strain is Mn1-7(T) ( = DSM 26584(T) = CCTCC AB 2012916(T)).

Response of Saccharomyces cerevisiae var. diastaticus to nerol: Evaluation of antifungal potential by inhibitory effect and proteome analyses.

Saccharomyces cerevisiae var. diastaticus (S. diastaticus) is a major spoilage yeast in brewing. In the present research, the antifungal properties of nerol and the proteome response of S. diastaticus were studied. Results showed nerol can inhibit cell budding and delay yeast fermentation in a dose-depended manner. After 3 d of treatment with 0.25 mg·mL-1 nerol, intracellular ROS levels increased 1.66-fold (P < 0.01), and the cells with damaged membrane increased to 23.2 %. Quantitative proteomic profiles utilizing a capillary-HPLC-MS/MS technology revealed that proteins involved in the metabolism of fermentable sugars were up-regulated in S. diastaticus cells treated with nerol, indicating nerol treatment altered the metabolite pattern of fermentable sugars. Proteins associated with the cell membrane biogenesis, heat shock proteins, amino acid biosynthesis, and glutathione metabolism were similarly up-regulated. These findings revealed the mechanism of nerol-induced yeast cell damage as well as the detoxification response of yeast cells.

Twelve natural estrogens and ten bisphenol analogues in eight drinking water treatment plants: Analytical method, their occurrence and risk evaluation.

Bisphenol analogues (BPs) and natural estrogens (NEs) as two important groups of endocrine-disrupting compounds (EDCs) in drinking water treatment plants (DWTPs) have been hardly investigated except bisphenol A (BPA) and three major NEs including estrone (E1), 17ß-estradiol (E2) and estriol (E3). In this study, a GC-MS analytical method was firstly established and validated for trace simultaneous determination of ten BPs and twelve NEs in drinking water, which included BPA, bisphenol B (BPB), bisphenol C (BPC), bisphenol E (BPE), bsiphenol F (BPF), bsiphenol P (BPP), bisphenol S (BPS), bisphenol Z (BPZ), bisphenol AF (BPAF), bisphenol AP (BPAP), E1, E2, E3, 17α-estradiol (17α-E2), 2-hydroestrone (2OHE1), 16hydroxyestrone (16α-OHE1), 4-hydroestrone (4OHE1), 2-hydroxyesstradiol (2OHE2), 4-hydroxyestradiol (4OHE2), 17-epiestriol (17epiE3), 16-epiestriol (16epiE3) and 16keto-estraiol (16ketoE2). This investigation showed that eighteen out of twenty-two targeted compounds were detected in drinking source waters of eight DWTPs with concentrations ranging from not detected to 142.8 ng/L. Although the conventional treatment process of DWTP could efficiently remove both BPs and NEs with respective removal efficiencies of 74.1%-90.9% and 74.5%-100%, BPA, BPS, BPE, BPZ, E1, 2OHE1, and 2OHE2 were found in the finished drinking waters. Chlorination could remove part of BPs and NEs, but the efficiency varied greatly with DWTP and the reason was unknown. In the finished drinking waters of eight DWTPs, the highest chemically calculated estrogen equivalence (EEQ) derived from BPs and NEs was up to 6.11 ngE2/L, which was over 22 times that could do harm to zebrafish, indicating a potential risk to human health. Given the fact that many chlorination products of BPs and NEs likely have higher estrogenic activities, the estrogenic effect of BPs and NEs in finished drinking water should be accurately examined urgently with the inclusion of BPs, NEs as well as their main chlorinated by-products. This study shed new light on the occurrence, removal, and potential estrogenic effects of BPs and NEs in DWTPs.

Simultaneous determination of twelve natural estrogens in dairy milk using liquid-liquid extraction and solid-phase extraction coupled with gas chromatography-mass spectrometry.

There have been many analytical methods for natural estrogens in commercial dairy milk samples, but in most of which, only four major estrogens (estrone (E1), 17ß-estradiol (E2), estriol (E3), and 17α-estradiol (αE2)) were included. This work developed an effective GC-MS analytical method for simultaneous analysis of twelve natural estrogens in commercial dairy milk sample, in which eight far-less well-known natural estrogens (2-hydroxyestone (2OHE1), 4-hydroxyestrone (4OHE1), 2-hydroxyestradiol (2OHE2), 4-hydroxyestradiol (4OHE2), 16-epiestriol (16epiE3), 16α-hydroxyestrone (16αOHE1), 16-ketoestradiol (16ketoE2) and 17epiestriol (17epiE3)) were included besides the four major natural estrogens. With liquid-liquid extraction and solid phase extraction, twelve natural estrogens in commercial dairy milk could be effectively extracted. The established method showed good linearity (R2 > 0.9991), low limits of detections (LODs, 0.02-0.11 ng/g), as well as excellent recoveries (64-117%) with satisfactory low relative standard deviations (RSDs, 0.8-14.7%). This established method was applied to seven commercial dairy milk samples, and all the twelve natural estrogens were frequently detected except for 4OHE2 without detection in any sample. Our results showed that the concentration contribution ratios of the eight far-less well-known natural estrogens in commercial dairy milk samples contributed to 32-83%, while the corresponding contribution ratios based on estrogen equivalence (EEQ) were 21-62%. This work highlighted the high abundance of the eight far-less well-known natural estrogens in commercial dairy milk based on both concentration and EEQ, which has been neglected for a long time.

Evolutionary rate of human tissue-specific genes are related with transposable element insertions.

The influence of transposable elements (TEs) on genome evolution has been widely studied. However, it remains unclear whether TE insertions also impact on evolutionary rate of human genes. In this study, we have compared the differences in TEs and evolutionary rates between human tissue-specific genes. Our results showed that various functional categories of human tissue-specific genes contained different TE numbers and divergent values of Ka/Ks, with human nucleic acid binding transcription factor activity genes having the fewest TE density and Ka/Ks value. Interestingly, we also found that human tissue-specific genes with TEs have also undergone faster evolution than those without TEs. Therefore, TEs have significant impact on the evolutionary rates of human tissue-specific genes. Furthermore, local genomic properties such as gene length, GC content and recombination rate may reflect a true transpositional bias for the particular TEs. Our results may provide important insights for further elucidating the evolution of human tissue-specific genes.

SLC41A1 Mg(2+) transport is regulated via Mg(2+)-dependent endosomal recycling through its N-terminal cytoplasmic domain.

SLC41A1 (solute carrier family 41, member A1) is a recently described vertebrate member of the MgtE family of Mg(2+) transporters. Although MgtE transporters are found in both prokaryotic and eukaryotic organisms, and are highly conserved, little is known about the regulation of their Mg(2+) transport function. In the present study, we have shown that endogenous SLC41A1 transporter expression is post-transcriptionally regulated by extracellular Mg(2+) in TRPM7 (transient receptor potential cation channel, subfamily M, member 7)-deficient cells, suggesting that SLC41A1 transporters underlie a novel plasma membrane Mg(2+) transport function. Consistent with this conclusion, structure-function analyses of heterologous SLC41A1 transporter expression demonstrate that SLC41A1 transporters exhibit the same plasma membrane orientation as homologous bacterial MgtE proteins, are capable of complementing growth of TRPM7-deficient cells only when the Mg(2+) transporting pore is intact, and require an N-terminal cytoplasmic domain for Mg(2+)-dependent regulation of lysosomal degradation and surface expression. Taken together, our results indicate that SLC41A1 proteins are a central component of vertebrate Mg(2+) transport systems, and that their Mg(2+) transport function is regulated primarily through an endosomal recycling mechanism involving the SLC41A1 N-terminal cytoplasmic domain.

RETRACTED ARTICLE: Down-regulation of lncRNA FEZF1-AS1 mediates regulatory T cell differentiation and further blocks immune escape in colon cancer.

Statement of RetractionWe, the Editors and Publisher of the journal Expert Review of Molecular Diagnostics, have retracted the following article:Sen Hong, Zhenkun Yan, YuMei Song, MiaoMiao Bi & Shiquan Li. Down-regulation of lncRNA FEZF1-AS1 mediates regulatory T cell differentiation and further blocks immune escape in colon cancer. Expert Review of Molecular Diagnostics. 2021. DOI: 10.1080/14737159.2022.2012157Since publication, significant concerns have been raised about the integrity of the data and reported results in the article. When approached for an explanation, the authors did not provide their original data or any necessary supporting information. As verifying the validity of published work is core to the integrity of the scholarly record, we are therefore retracting the article. The corresponding author listed in this publication has been informed.We have been informed in our decision-making by our policy on publishing ethics and integrity and the COPE guidelines on retractions.The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as 'Retracted'.

Exploring two types of prenylated bitter compounds from hop plant (Humulus lupulus L.) against α-glucosidase in vitro and in silico.

Hops are abundant in natural bioactive compounds. In this work, nine prenylated bitter compounds from hop were evaluated for their inhibitory activity against α-glucosidase. As a result, four flavonoids and one phloroglucinol (lupulone, LP) outperformed acarbose in inhibiting α-glucosidase. Isoxanthohumol (IX) and LP with two types of structures were selected for inhibition mechanism studies by spectroscopic methods and molecular dynamics simulation (MD). Results showed that IX acted as noncompetitive inhibitor and bound to α-glucosidase in allosteric sites via hydrogen bonds, hydrophobic, van der Waals (vdW), and electrostatic force, whereas LP was uncompetitive inhibitor and bound to catalytic sites via hydrophobic and vdW interactions. Notably, the conformation around binding site of α-glucosidase formed stable α-helix and tightened after binding IX and LP, respectively, which helped to elucidate noncompetitive and uncompetitive inhibitory mechanisms. This work demonstrated that two types of prenylated bitter compounds are discrepant in their mechanisms of interaction with α-glucosidase.

microR-4449 Promotes Colorectal Cancer Cell Proliferation via Regulation of SOCS3 and Activation of STAT3 Signaling.

INTRODUCTION: Dysregulation of microRNAs (miRNAs), which represented a critical level of gene expression modulation, regulated the development of colorectal cancer. However, the functions of numerous miRNAs remain unclear in colorectal cancer. METHODS: The microarray data of GSE115513 were retrieved; subsequently, the differentially expressed miRNAs between 411 colon tumors and 381 normal colon mucosa were analyzed. Real-time PCR (RT-qPCR) and bioinformatic analysis were applied to examine the expression of miR-4449 in collected colorectal tumors and published microarray data. The activity of signal transducer and activator of transcription 3 (STAT3) signaling was detected by Western blotting and RT-qPCR. Dual-Luciferase assay and bioinformatic analysis were used to confirm the interaction between suppressor of cytokine signaling 3 (SOCS3) and miR-4449. Loss of function and rescue assays were performed to study the involvement of miR-4449 and SOCS3 in cell proliferation and apoptosis of colorectal cancer. RESULTS: Herein, we identified miR-4449 as a novel upregulated miRNA in colorectal cancer. Our data suggested that miR-4449 downregulation blocked the proliferation of colorectal cancer cells accompanied with the elevation of cell apoptosis. Decreased expression of miR-4449 led to inactivation of STAT3 pathway as indicated by dephosphorylation of STAT3 and downregulation of STAT3 target genes, including vascular endothelial growth factor (VEGF), c-Myc, baculovirus inhibitor of apoptosis containing 5 (BIRC5). Furthermore, SOCS3, a negative regulator of STAT3 pathway, was found to be a target gene of miR-4449. The data also showed that the inactivation of STAT3 pathway by miR-4449 inhibitor was realized by targeting SOCS3. Moreover, the biological function of miR-4449 downregulation was reversed by SOCS3 knockdown in colorectal cancer cells. CONCLUSION: The current study revealed that miR-4449 promoted cell proliferation of colorectal cancer and was a promising potential therapeutic target for colorectal cancer.

Combined effect of microplastics and DDT on microbial growth: A bacteriological and metabolomics investigation in Escherichia coli.

Microplastics (MPs) can adsorb toxic chemicals in biological or environmental matrixes and thus influence their behavior and availability. In order to investigate how the combined pollution of MPs and toxic organic chemical influence microbial growth and metabolism, Escherichia coli (E. coli) was grown in a complex, well-defined media and treated with polystyrene microplastics (PS MPs) and dichloro-diphenyl-tricgloroethane (DDT) at human relevant concentration levels. In vivo metabolites captured by a novel solid phase microextraction (SPME) probe, were used to reflect the metabolic dysregulation of E. coli under different pollution stresses. Results showed that the toxic effect of DDT displayed a distinct dose-dependent phenomenon while the existence of PS decreased the growth and metabolic interference effect of DDT on E. coli. Adsorption results revealed a mechanism that PS weakened the adverse impact of DDT by decreasing its free concentration in the treated culture media. Tricarboxylic acid (TCA) cycle related enzymes activities and antioxidant defense related substances of E. coli also proved the mechanism. The current study is believed to broaden our understanding of the ecotoxicity of MPs with toxic organic chemicals on microorganism.

Distribution and potential provenance of trace elements in a 120-year dated sediment core from west Daya Bay, northeastern South China Sea.

Eighteen trace elements were analyzed in a 120-year sediment core from Daya Bay. Burial flux history and potential provenance, the relationships among trace elements, and biogenic compositions were analyzed for determining the trend and extent of trace element accumulation and identifying corresponding anthropogenic effects. Additionally, the effects of anthropogenic activities on Daya Bay were reconstructed, and a baseline/background estimation was provided for Daya Bay. The burial fluxes of V, Cr, Cd, Cu, Zn, Mn, Fe, Co, Ni, Pb, Hg, Zn, Mo, Ag, As, Se, and Tl increased from 1960 to 2010, especially after the late 1980s. Our results are useful for understanding pollution and land-sea interactions along the coasts of the South China Sea, especially in the Guangdong-Hong Kong-Macao Greater Bay Area.

A novel signaling pathway: fibroblast nicotinic receptor alpha1 binds urokinase and promotes renal fibrosis.

The nicotinic acetylcholine receptor alpha1 (nAChRalpha1) was investigated as a potential fibrogenic molecule in the kidney, given reports that it may be an alternative urokinase (urokinase plasminogen activator; uPA) receptor in addition to the classical receptor uPAR. In a mouse obstructive uropathy model of chronic kidney disease, interstitial fibroblasts were identified as the primary cell type that bears nAChRalpha1 during fibrogenesis. Silencing of the nAChRalpha1 gene led to significantly fewer interstitial alphaSMA(+) myofibroblasts (2.8 times decreased), reduced interstitial cell proliferation (2.6 times decreased), better tubular cell preservation (E-cadherin 14 times increased), and reduced fibrosis severity (24% decrease in total collagen). The myofibroblast-inhibiting effect of nAChRalpha1 silencing in uPA-sufficient mice disappeared in uPA-null mice, suggesting that a uPA-dependent fibroblastic nAChRalpha1 pathway promotes renal fibrosis. To further establish this possible ligand-receptor relationship and to identify downstream signaling pathways, in vitro studies were performed using primary cultures of renal fibroblasts. (35)S-Labeled uPA bound to nAChRalpha1 with a K(d) of 1.6 x 10(-8) m, which was displaced by the specific nAChRalpha1 inhibitor d-tubocurarine in a dose-dependent manner. Pre-exposure of uPA to the fibroblasts inhibited [(3)H]nicotine binding. The uPA binding induced a cellular calcium influx and an inward membrane current that was entirely prevented by d-tubocurarine preincubation or nAChRalpha1 silencing. By mass spectrometry phosphoproteome analyses, uPA stimulation phosphorylated nAChRalpha1 and a complex of signaling proteins, including calcium-binding proteins, cytoskeletal proteins, and a nucleoprotein. This signaling pathway appears to regulate the expression of a group of genes that transform renal fibroblasts into more active myofibroblasts characterized by enhanced proliferation and contractility. This new fibrosis-promoting pathway may also be relevant to disorders that extend beyond chronic kidney disease.

LncRNA AGAP2-AS1 augments cell viability and mobility, and confers gemcitabine resistance by inhibiting miR-497 in colorectal cancer.

BACKGROUND: Most recently, long non-coding RNAs (lncRNAs) emerge as crucial modulators in many biological processes, such as embryonic development, cell growth, and tumorigenesis. However, the correlations between lncRNAs and colorectal cancer (CRC) cell proliferation, metastasis, and gemcitabine resistance are not well understood. RESULTS: The expression of AGAP2-AS1 was overexpressed in CRC tissues and negatively correlated with the survival of patients with CRC. AGAP2-AS1 promoted CRC cell proliferation and inhibited apoptosis. Moreover, AGAP2-AS1 enhanced the chemoresistance of CRC cells to gemcitabine. In addition, AGAP2-AS1 enhanced the migration and invasion of CRC cells. Mechanistic studies showed that AGAP2-AS1 regulated fibroblast growth factor receptor 1 (FGFR1) expression by sponging miR-497 in CRC progression. CONCLUSION: We identified an oncogenic role of AGAP2-AS1 in the development and progression of CRC. METHODS: qRT-PCR was used to measure the expression of AGAP2 Antisense RNA 1 (AGAP2-AS1) in 116 cases of CRC and adjacent normal tissues. Luciferase reporter assays was used to detect the interaction between AGAP2-AS1 and miR-497. The xenograft tumor experiment was used to study the in vivo function of AGAP2-AS1.

LncRNA TUG1 promotes the progression of colorectal cancer via the miR-138-5p/ZEB2 axis.

To explore the role of long-chain non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in the development of colorectal cancer (CRC) via the miR-138-5p/zinc finger E-box-binding homeobox 2 (ZEB2) axis. Eighty-four CRC tissue specimens and 84 corresponding paracancerous tissue specimens were sampled from 84 patients with CRC admitted to the First Hospital of Jilin University from January 2018 to September 2019. The TUG1 expression in the specimens was determined, and its value in diagnosis and prognosis of CRC was analyzed. Additionally, constructed stable and transient overexpresison vectors and inhibition vectors were transfected into CRC cells. The MTT, transwell, and flow cytometry were adopted for analysis on the proliferation, invasion, and apoptosis of transfected cells, respectively, and a dual luciferase reporter (DLR) assay was carried out for correlation determination between TUG1 and miR-138-5p and between miR-138-5p and ZEB2. TUG1 was up-regulated in CRC, and serum TUG1 could be adopted as a diagnostic marker of CRC, with area-under-the-curve (AUC) larger than 0.8. In addition, siRNA-TUG1, shRNA-TUG1, miR-138-5p-mimics, and miR-138-5p-inhibitor were transfected into cells, and it turned out that overexpressing miR-138-5p and inhibiting ZEB2 exerted the same effects. The DLR assay revealed that TUG1 was able to targetedly regulate miR-138-5p, and miR-138-5p could targetedly regulate ZEB2, and in vitro experiments revealed that TUG1 could affect the epithelial-to-mesenchymal transition (EMT) of CRC via the miR-138-5p/ZEB2 axis. TUG1 could promote the development of CRC via the miR-138-5p/ZEB2 axis.

miR-663b promotes colorectal cancer progression by activating Ras/Raf signaling through downregulation of TNK1.

MiR-663b has been demonstrated to be abnormally expressed in several cancer types and was involved in the progression of cancer. Although overexpression of miR-663b in colorectal cancer was observed, the role of miR-663b in colorectal cancer cells has not been identified yet. In this study, we analyzed expression of miR-663b in colorectal tumors and explored the molecular mechanism of miR-663b in colorectal cancer cells. MiR-663b was significantly overexpressed in colorectal tumors and cell lines. Downregulation of miR-663b inhibited cell proliferation and sphere forming ability in colorectal cancer cells. In addition, miR-663b downregulation inactivated Ras/Raf signaling activity and subsequently decreased YAP1 and CD44 expression in colorectal cancer cells. Using TargetScan software, TNK1, a negative regulator of Ras/Raf signaling, was predicted to be a target gene of miR-663b. Western blotting and RT-qPCR showed that TNK1 expression was negatively regulated by miR-663b. In addition, the direct binding of miR-663b to TNK1 mRNA was proved by dual luciferase reporter assay. Furthermore, downregulation of miR-663b inhibited colorectal cancer cell proliferation and stemness, which was reversed after siRNA-mediated silencing of TNK1. In conclusion, the current study revealed a pivotal role of miR-663b in the progression of colorectal cancer.