Identification and Validation of a Novel Tertiary Lymphoid Structures-Related Prognostic Gene Signature in Hepatocellular Carcinoma.

Background: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors originating from the digestive system. Tertiary lymphoid structures (TLS), non-lymphoid tissues outside of the lymphoid organs, are closely connected to chronic inflammation and tumorigenesis. However, the detailed relationship between TLS and HCC prognosis remained unclear. In this study, we aimed to construct a TLS-related gene signature for predicting the prognosis of HCC patients. Methods: The Cancer Genome Atlas (TCGA) clinical data from 369 HCC tissues and 50 normal liver tissues were utilized to examine the differential expression of TLS-related genes. Based on least absolute shrinkage and selection operator (LASSO) Cox regression analysis, the prognostic model was constructed using the TCGA cohort and validated in the GSE14520 cohort and International Cancer Genome Consortium (ICGC) cohort. The Kaplan-Meier (KM) and receiver operating characteristic (ROC) curves were employed to validate the predictive ability of the prognostic model. Furthermore, Cox regression analysis was applied to identify whether the TLS score could be employed as an independent prognosis factor. A nomogram was developed to predict the survival probability of HCC patients. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were performed for TLS-related genes. Genetic mutation analysis, the CIBERSORT algorithm, and single-sample gene set enrichment analysis (ssGSEA) were used to assess the tumor mutation landscape and immune infiltration. Finally, the role of the TLS score in HCC therapy was investigated. Results: Six genes were included in the construction of our prognostic model (CETP, DNASE1L3, PLAC8, SKAP1, C7, and VNN2), and we validated its accuracy. Survival analysis showed that patients in the high-TLS score group had a significantly better overall survival than those in the low-TLS score group. Univariate, multivariate Cox regression analysis and the establishment of a nomogram indicated that the TLS score could independently function as a potential prognostic marker. A significant association between TLS score and immunity was revealed by an analysis of gene alterations and immune cell infiltration. In addition, two subtypes of the TLS score could accurately predict the effectiveness of sorafenib, transcatheter arterial chemoembolization (TACE), and immunotherapy in HCC patients. Conclusion: In this research, we conducted and validated a prognostic model associated with TLS that may be helpful for predicting clinical outcomes and treatment responsiveness for HCC patients.

Protective effect of scallop-derived plasmalogen against vascular dysfunction, via the pSTAT3/PIM1/NFATc1 axis, in a novel mouse model of Alzheimer's disease with cerebral hypoperfusion.

A strong relationship between Alzheimer's disease (AD) and vascular dysfunction has been the focus of increasing attention in aging societies. In the present study, we examined the long-term effect of scallop-derived plasmalogen (sPlas) on vascular remodeling-related proteins in the brain of an AD with cerebral hypoperfusion (HP) mouse model. We demonstrated, for the first time, that cerebral HP activated the axis of the receptor for advanced glycation endproducts (RAGE)/phosphorylated signal transducer and activator of transcription 3 (pSTAT3)/provirus integration site for Moloney murine leukemia virus 1 (PIM1)/nuclear factor of activated T cells 1 (NFATc1), accounting for such cerebral vascular remodeling. Moreover, we also found that cerebral HP accelerated pSTAT3-mediated astrogliosis and activation of the nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) inflammasome, probably leading to cognitive decline. On the other hand, sPlas treatment attenuated the activation of the pSTAT3/PIM1/NFATc1 axis independent of RAGE and significantly suppressed NLRP3 inflammasome activation, demonstrating the beneficial effect on AD.

Neferine exerts anti­inflammatory activity in BV­2 microglial cells and protects mice with MPTP­induced Parkinson's disease by inhibiting NF­κB activation.

Abnormal activation of microglia and the production of proinflammatory cytokines can lead to chronic neuroinflammation, which is an important pathological characteristic of Parkinson's disease (PD). Neferine is a chemical compound extracted from lotus seed which has previously been reported to exert protective effects on the development of several types of cancer, myocardial injury and hypoxic­ischemic encephalopathy. However, its effect on microglial functions in neuroinflammation remains to be clarified. The present study used network pharmacology and screening in a lipopolysaccharide (LPS) model to demonstrate that neferine suppresses the production of inducible nitric oxide synthase, interleukin­6 and tumor necrosis factor α in LPS­treated BV­2 cells. The working concentration of neferine did not exert cytotoxic effects on BV­2 cells. Mechanistically, neferine attenuated inflammation by inhibiting the phosphorylation and nuclear translocation of the NF­κB p65 subunit. In vivo, neferine protected mice from the inflammatory response in the substantia nigra and inhibited the development of nervous disorders in the 1­methyl­4­phenyl­1,2,3,6­tetrahydropyridine­induced PD model. The present study demonstrated that neferine inhibited LPS­mediated activation of microglia by inhibiting NF­κB signaling. These findings may provide a new reference for the prevention and future treatment of PD.

Intrinsic Molecular Mechanisms of Transformation between Isomeric Intermediates Formed at Different Stages of Cysteine-Xylose Maillard Reaction Model through Dehydration.

2-Threityl-thiazolidine-4-carboxylic acid (TTCA) and Amadori rearrangement product (ARP), the isomeric intermediates derived from the cysteine-xylose (Cys-Xyl) Maillard reaction model, possessed the ability to produce similar flavor profile during the thermal process, but the flavor formation or browning rate of heated TTCA was significantly lower than that of ARP. Macroscopically, the yield of TTCA reached the maximum when the moisture content of the reaction system just dropped to nearly 0% during the thermal reaction-vacuum dehydration process. During the subsequent dynamic intramolecular dehydration process, the reaction remained at an early stage of the Maillard reaction, and TTCA was the main intermediate. Thereinto, the water activity of the samples decreased with the increased dehydration time. From a molecular perspective, the dissipation of free water promoted the conversion of combined water to immobilized water and free water, increasing the intramolecular dehydration. Instantaneous high-temperature dehydration during the spray drying process revealed a higher efficiency than the thermal reaction-vacuum dehydration process, which facilitated the specific conversion of substrates to intermediates (TTCA, ARP). The loss of free water and immobilized water was a key driving force for the direct formation of TTCA/ARP, regulating the formation stages of MRIs. The increase of the inlet air temperature could alter the ratio of TTCA and ARP at the equilibrium state.

Regulated Formation of Inhibited Color and Enhanced Flavor Derived from Heated 2-Threityl-Thiazolidine-4-Carboxylic Acid with Additional Cysteine Targeting at Different Degradation Stages.

This study explored the addition of cysteine (Cys) affecting the color formation of heated 2-threityl-thiazolidine-4-carboxylic acid (TTCA) models under different reaction conditions and pointed out that temperature was considered to be the key parameter influencing the color inhibition behavior of Cys on TTCA reaction models. Results revealed that additional Cys not only controlled the reaction progress and blocked the formation pathway of browning but also changed the formation rate, intensity, and profile of the flavor generated from the TTCA reaction model. Meanwhile, the mechanism of Cys simultaneously regulating the formation of color and flavor was revealed through monitoring of the characteristic downstream products during TTCA degradation and model reaction systems. At the initial stage, the additional Cys acted as a color inhibitor before the deoxyxylosone degradation, preventing the formation of downstream browning precursors. With the continuous depletion of Cys as well as the generation of furans or α-dicarbonyl compounds, Cys became a flavor enhancer to act on the browning precursors and to provide more sulfur/nitrogen elements for the TTCA thermal reaction system. Therefore, Cys had the potential to act as both color inhibitor and flavor fortifier to match with TTCA for the preparation of a light-colored flavoring base with a desired flavor during thermal processing.

miR-424-3p promotes metastasis of hepatocellular carcinoma via targeting the SRF-STAT1/2 axis.

Although emerging evidence has established the roles of miRNAs in hepatocellular carcinoma (HCC), the global functional implication of miRNAs in this malignancy remains largely uncharacterized. Here, we aim to systematically identify novel miRNAs involved in HCC and clarify the function and mechanism of specific novel candidate miRNA(s) in this malignancy. Through an integrative omics approach, we identified ten HCC-associated functional modules and a collection of candidate miRNAs. Among them, we demonstrated that miR-424-3p, exhibiting strong associations with extracellular matrix (ECM), promotes HCC cells migration and invasion in vitro and facilitates HCC metastasis in vivo. We further demonstrated that SRF is a direct functional target of miR-424-3p, and is required for the oncogenic activity of miR-424-3p. Finally, we found that miR-424-3p reduces the interferon pathway by attenuating the transactivation of SRF on STAT1/2 and IRF9 genes, which in turn enhances the matrix metalloproteinases (MMPs)-mediated ECM remodeling. This study provides comprehensive functional relevance of miRNAs in HCC by an integrative omics analysis, and further clarifies that miR-424-3p in ECM functional module plays an oncogenic role via reducing the SRF-STAT1/2 axis in this malignancy.

Prevalence and risk factors for dementia in the Tibetan region: A population-based cross-sectional study.

BACKGROUND: As China's population ages, the nationwide prevalence of dementia is increasing. However, the epidemiology of dementia among the Tibetan population remains unclear. OBJECTIVE: A cross-sectional study was conducted involving 9116 participants aged >50 years in the Tibetan population to investigate the risk factors and prevalence of dementia among this population. Permanent residents of the region were invited to participate, and the response rate was 90.7 %. METHODS: The participants underwent neuropsychological testing and clinical assessments, from which physical measurements (e.g., body mass index, blood pressure), demographic information (e.g., gender, age), and lifestyle details (e.g., family living arrangement, smoking, alcohol arrangement) were recorded. Dementia diagnoses were made using the standard consensus diagnostic criteria. The risk factors of dementia were identified using stepwise multiple logistic regression. RESULTS: The average age of the participants was 63.71 (standard deviation = 9.36), and there were 44.86 % males. The prevalence of dementia was 4.66 %. The multivariate logistic regression analysis revealed that older age, unmarried status, lower education level, obesity, hypertension, diabetes, coronary heart disease, cerebral vascular disease, and HAPC were independently and positively associated with dementia (P < 0.05). However, no association was found between the frequency of religious activities and the prevalence of dementia in this population (P > 0.05). CONCLUSIONS: There exist a number of contributory risk factors for dementia in the Tibetan population, with variations associated with high altitude, religious activities (i.e., scripture turning, chanting, spinning Buddhist beads, and bowing), and dietary habits. These findings suggest that social activities, such as religious activities, are protective factors for dementia.

Cysteine-Induced pH-Dependent Formation of Thiols and Sulfides or 2-Acetylthiazole and Pyrazines during Thermal Treatment of N-(1-Deoxy-d-xylulos-1-yl)-alanine.

The influence of pH was studied on volatile flavor formation during thermal treatment of an Amadori rearrangement product (ARP) with or without the addition of cysteine (Cys). The formation of thiols and sulfides or 2-acetylthiazole and pyrazines induced by Cys during thermal degradation of ARP was pH-dependent. At low pH levels, the hydrolysis of Cys to hydrogen sulfide (H2S) was promoted, giving rise to the increase of thiols and sulfides with an obvious meaty aroma. However, alkaline conditions were beneficial for enhancing the cyclization or transformation of imine to the enol structure, which strengthened the formation of 2-acetylthiazole and pyrazines with a roasted and nutty aroma. The imine was derived from the nucleophilic addition of Cys and methylglyoxal (MGO) and subsequent decarboxylation. At pH 8, Cys-induced variation of the flavor profile was weakened during thermal degradation of ARP. Accordingly, the combinational effect of pH and added Cys could be beneficial for achieving the desirable flavors during thermal processing of ARP.

Reduced asynchronism between regenerative cysteine and fragments of deoxyosones promoting formation of sulfur-containing compounds through extra-added xylose and elevated temperature during thermal processing of 2­threityl-thiazolidine-4-carboxylic acid.

The flavor intensity of thermally processed 2­threityl-thiazolidine-4-carboxylic acid (TTCA) was significantly improved to 1.56 times of that generated from MRPs, but its flavor profile was not as desirable as that of fresh MRPs. The synergistic effect between the additional xylose (Xyl) and elevated temperature was proposed and confirmed via the quantitative analyses of regenerative cysteine (Cys) and fragments of deoxyosones (MGO/GO), which reduced the asynchronism between the formation of released Cys from degraded TTCA and retro-aldolisation products of the intermediate deoxyosones. This synergistic effect further enhanced the Strecker degradation of Cys as well as its thermal degradation and thereby promoted the formation of characteristic flavor substances including sulfur-containing compounds and pyrazines, and the total concentrations of TTCA reaction model reached 205.954 µg/L with additional Xyl at 140 °C. Model reaction systems were employed to verify this hypothesis and the proposed mechanism was further elucidated through isotope labeling technique.

The A-to-I editing of KPC1 promotes intrahepatic cholangiocarcinoma by attenuating proteasomal processing of NF-κB1 p105 to p50.

BACKGROUND: Aberrant RNA editing of adenosine-to-inosine (A-to-I) has been linked to multiple human cancers, but its role in intrahepatic cholangiocarcinoma (iCCA) remains unknown. We conducted an exome-wide investigation to search for dysregulated RNA editing that drive iCCA pathogenesis. METHODS: An integrative whole-exome and transcriptome sequencing analysis was performed to elucidate the RNA editing landscape in iCCAs. Putative RNA editing sites were validated by Sanger sequencing. In vitro and in vivo experiments were used to assess the effects of an exemplary target gene Kip1 ubiquitination-promoting complex 1 (KPC1) and its editing on iCCA cells growth and metastasis. Crosstalk between KPC1 RNA editing and NF-κB signaling was analyzed by molecular methods. RESULTS: Through integrative omics analyses, we revealed an adenosine deaminases acting on RNA 1A (ADAR1)-mediated over-editing pattern in iCCAs. ADAR1 is frequently amplified and overexpressed in iCCAs and plays oncogenic roles. Notably, we identified a novel ADAR1-mediated A-to-I editing of KPC1 transcript, which results in substitution of methionine with valine at residue 8 (p.M8V). KPC1 p.M8V editing confers loss-of-function phenotypes through blunting the tumor-suppressive role of wild-type KPC1. Mechanistically, KPC1 p.M8V weakens the affinity of KPC1 to its substrate NF-κB1 p105, thereby reducing the ubiquitinating and proteasomal processing of p105 to p50, which in turn enhances the activity of oncogenic NF-κB signaling. CONCLUSIONS: Our findings established that amplification-driven ADAR1 overexpression results in overediting of KPC1 p.M8V in iCCAs, leading to progression via activation of the NF-κB signaling pathway, and suggested ADAR1-KPC1-NF-κB axis as a potential therapeutic target for iCCA.

Glycine, Diglycine, and Triglycine Exhibit Different Reactivities in the Formation and Degradation of Amadori Compounds.

A series of Amadori compounds of glucose were prepared from glycine (G-ARP), diglycine (DiG-ARP), and triglycine (TriG-ARP), and identified by UPLC-MS/MS and NMR. The formation rate of ARPs was TriG-ARP > DiG-ARP > G-ARP, and their activation energies were 63.48 kJ/mol (TriG-ARP), 72.84 kJ/mol (DiG-ARP), and 84.76 kJ/mol (G-ARP), respectively, suggesting that ARP was formed more easily from small peptides than from amino acid. Although 1-DG was formed much more difficultly than 3-DG, the same order of the formation of 1-DG, 3-DG, and browning was DiGly > TriGly > Gly. It was also confirmed that more methylglyoxal and glyoxal would be formed from small peptides than equimolar amino acids. Compared with free amino acid, ARP, deoxyglycosones, and their secondary degradation products were more easily formed from dipeptide and tripeptide, thereby stronger browning occurred and higher reactivity was exhibited in Maillard reaction of di- or tripeptide.

Degradation of 2-Threityl-Thiazolidine-4-Carboxylic Acid and Corresponding Browning Accelerated by Trapping Reaction between Extra-Added Xylose and Released Cysteine during Maillard Reaction.

2-Threityl-thiazolidine-4-carboxylic acid (TTCA), a nonvolatile precursor of flavor and color, is considered to be more stable than its isomeric Amadori compound (ARP). The degradation behavior of TTCA favors higher temperatures and pH. In order to adjust and control the thermal degradation of TTCA to improve its food processing adaptability, a TTCA-Xyl thermal reaction model was constructed to explore the effect of extra-added Xyl on the thermal degradation behavior of TTCA. The results confirmed that the extra-added Xyl was involved in the degradation pathway of TTCA and accelerated its depletion, thus promoting the formation of characteristic downstream products of TTCA including some α-dicarbonyl compounds, and consequently accelerating the browning formation. The isotope-labeling technique was further applied to confirm that the added Xyl could trap the Cys released from the decomposition of ARP and formed additional TTCA, which could promote the movement of chemical equilibrium and gradually accelerate the degradation rate of TTCA as well as melanoidins formation. The higher pH value could even promote this phenomenon.

An innovative adjustable prone positioning frame for treatment of severe kyphosis secondary to ankylosing spondylitis with two-level osteotomy.

PURPOSE: This study aims to introduce an innovative adjustable prone positioning frame (APPF) and explore its feasibility and safety for treatment of severe kyphosis secondary to ankylosing spondylitis (AS) with two-level osteotomy. METHODS: A retrospective, non-controlled study was conducted to illustrate the process where 13 patients diagnosed with severe kyphosis secondary to AS received operations on the APPF. Parameters of chin brow vertical angle (CBVA), global kyphosis (GK), thoracolumbar kyphosis (TLK), lumbar lordosis (LL) and sagittal vertical axis (SVA) were measured. Positioning time, operation time, intraoperative blood loss ahd complications were also determined. The Scoliosis Research Society outcomes instrument (SRS-22) was applied for clinical assessment. RESULTS: All patients were placed on the APPF successfully with the positioning time of 2.92 ± 0.76 min, received operation with 457.00 ± 88.04 min and had blood loss of 2330.77 ± 1423.25 ml. Four cases experienced pain due to tensional skin of the abdomen and one case suffered cerebrospinal fluid leakage postoperatively, but these patients were all cured conservatively. No neurological complications were observed, although sagittal translation occurred in four patients. Significant improvements were detected in CBVA, GK, TLK, LL and SVA postoperatively (P < 0.05), but no significant difference was observed between postoperation and the final follow-up (P > 0.05). The SRS-22 scores at 2 years after operation were significantly higher than those before operation (P < 0.05). CONCLUSION: The innovative APPF provided great convenience to place patients with severe kyphosis secondary to AS in a prone position. Performing two-level osteotomy with the aid of APPF is safe, feasible and effective.

The Human-Specific STING Agonist G10 Activates Type I Interferon and the NLRP3 Inflammasome in Porcine Cells.

Pigs have anatomical and physiological characteristics comparable to those in humans and, therefore, are a favorable model for immune function research. Interferons (IFNs) and inflammasomes have essential roles in the innate immune system. Here, we report that G10, a human-specific agonist of stimulator of interferon genes (STING), activates both type I IFN and the canonical NLRP3 inflammasome in a STING-dependent manner in porcine cells. Without a priming signal, G10 alone transcriptionally stimulated Sp1-dependent p65 expression, thus triggering activation of the nuclear factor-κB (NF-κB) signaling pathway and thereby priming inflammasome activation. G10 was also found to induce potassium efflux- and NLRP3/ASC/Caspase-1-dependent secretion of IL-1ß and IL-18. Pharmacological and genetic inhibition of NLRP3 inflammasomes increased G10-induced type I IFN expression, thereby preventing virus infection, suggesting negative regulation of the NLRP3 inflammasome in the IFN response in the context of STING-mediated innate immune activation. Overall, our findings reveal a new mechanism through which G10 activates the NLRP3 inflammasome in porcine cells and provide new insights into STING-mediated innate immunity in pigs compared with humans.

Transformation between 2-Threityl-thiazolidine-4-carboxylic Acid and Xylose-Cysteine Amadori Rearrangement Product Regulated by pH Adjustment during High-Temperature Instantaneous Dehydration.

2-Threityl-thiazolidine-4-carboxylic acid (TTCA) was found to be the predominant product rather than the Amadori rearrangement product (ARP) during the formation of xylose-cysteine-derived (Xyl-Cys-derived) Maillard reaction intermediates (MRIs) through a thermal reaction coupled with vacuum dehydration. To regulate the existence forms of Xyl-Cys-derived MRIs, an effective method carried out by pH adjustment during high-temperature instantaneous dehydration through spray-drying was proposed in this research to promote the conversion from TTCA to ARP. The increased inlet air temperature of spray-drying could properly facilitate the shift of chemical equilibrium between the MRIs and promote the transformation from TTCA to ARP while raising the total yield of TA (TTCA + ARP). The conversion to ARP was increased to 20.83% at 190 °C of hot blast compared to the product without spray-drying (6.03%). The conversion from TTCA to ARP was further facilitated in the pH range of 7.5-9.5. When the pH of the aqueous reactants was adjusted to 9.5, the equilibrium conversion to ARP was improved to 47.23% after spray-drying, which accounted for 59.48% of the TA formation. Accordingly, MRIs with different TTCA/ARP proportions could be selectively obtained by pH adjustment of the stock solution during high-temperature instantaneous dehydration of spray-drying.

Interaction of Added l-Cysteine with 2-Threityl-thiazolidine-4-carboxylic Acid Derived from the Xylose-Cysteine System Affecting Its Maillard Browning.

The Maillard reaction under a stepwise increase of temperature using l-cysteine as the indicator was performed to determine the formation conditions for the preparation of 2-threityl-thiazolidine-4-carboxylic acid (TTCA) which was the main Maillard reaction intermediate (MRI) derived from the xylose (Xyl)-cysteine (Cys) model system in aqueous medium. To clarify the indicating mechanism of Cys for the TTCA formation, the thermal model systems of TTCA-Cys and TTCA solutions were investigated. The browning of the final Maillard reaction products (MRPs) and concentration of downstream degradation products of MRIs indicated that the added Cys could react with TTCA to inhibit the formation of visible color via preventing the generation of dicarbonyl compounds derived from MRIs. Through HPLC analysis, it was demonstrated that added Cys affected the normal reaction pathway from TTCA to ARP and other downstream products by restoring TTCA to sugar and amino acid under heat treatment.

Ginkgetin aglycone attenuates neuroinflammation and neuronal injury in the rats with ischemic stroke by modulating STAT3/JAK2/SIRT1.

The present investigation evaluates the protective effect of Ginkgetin aglycone (GA) against ischemic stroke-induced neuronal injury. Ischemic stroke was produced by the middle cerebral artery occlusion (MCAO) model and animals were a group that received GA 100 and 200 mg/kg, i.p. five days before the induction of MCAO. The effect of GA against stroke was determined by estimating the neurological deficit score and brain water content was also observed. Moreover terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay was done for determining the neuronal apoptosis and Western blot assay also performed for estimating the expression of several proteins. Results of the study suggest that the neurological deficit score and brain water content was found to be lower in the GA treated group than the ischemia/reperfusion (I/R) group of rats. Moreover the number of TUNEL positive cells was found to be lower in the GA treated group than in the I/R group of rats. There was a significant (p < 0.01) decrease in the oxidative stress parameters and cytokine in the tissue homogenate of the GA treated group compared to the I/R group of rats. Further treatment with GA attenuates altered expression of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), protein kinase B (Akt), B-cell lymphoma 2 (Bcl-2), signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa light chain enhancer of activated B cells (NF-B), toll-like receptor 4 (TLR-4), Janus kinase 2 (JAK-2) and sirtuin-1 (SIRT-1) protein in the brain tissues of stroke rats. In conclusion, data of the report reveal that treatment with Ginkgetin aglycone protects the neuronal injury against stroke in rats by reducing oxidative stress and inflammation.

Association of MCP-1 promoter polymorphism with susceptibility to nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is prevalent among populations from southern China and is influenced by both genetic and environmental risk factors. The monocyte chemoattractant protein-1 (MCP-1), a member of cysteine-cysteine chemokine family, plays critical roles in cancers. A polymorphism within the MCP-1 promoter, rs1024611, has been shown to be significantly associated with the risk of several cancers. Our purpose was to assess the role of rs1024611 in NPC susceptibility. By polymerase chain reaction-restriction fragment length polymorphism method, we genotyped rs1024611 in 593 patients with NPC (cases) and 480 cancer-free subjects (controls) among Guangxi population from southern China. We observed that the G allele of rs1024611 was significantly associated with the increased risk of NPC in an additive model and dominant model, respectively (P = 0.018 and 0.010, odds ratio = 1.25 and 1.41, respectively). No appreciable variation of the effects was found across the subgroups stratified by age, sex, nationality, smoking and drinking status, and smoking level. In addition, significantly higher messenger RNA (mRNA) expression level of MCP-1 was observed in NPC tissues than that in normal nasopharyngeal tissues, and the G allele of rs1024611 was significantly associated with elevated mRNA expression level of MCP-1 in Epstein-Barr virus-transformed lymphocytes. In conclusion, our findings suggested that rs1024611 at the MCP-1 promoter may be a risk factor for NPC. Further studies with larger sample size are necessary to confirm these findings.

Germline Duplication of SNORA18L5 Increases Risk for HBV-related Hepatocellular Carcinoma by Altering Localization of Ribosomal Proteins and Decreasing Levels of p53.

BACKGROUND & AIMS: Single nucleotide polymorphisms could affect risk for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). We performed a germline copy number variation (CNV)-based genome-wide association study (GWAS) in populations of Chinese ancestry to search for germline CNVs that increase risk of HCC. METHODS: We conducted a CNV-based GWAS of 1583 HCC cases (persons with chronic HBV infection and HCC) and 1540 controls (persons with chronic HBV infection without HCC) in Chinese populations. Identified candidates were expressed in L-02, HepG2, or TP53-/- or wild-type HCT116 cells, and knocked down with short hairpin RNAs in HepG2, Bel-7402, and SMMC-7721 cells; proliferation, colony formation, and apoptosis were measured. Formation of xenograft tumors from cell lines was monitored in nude mice. Subcellular localization of ribosome proteins and levels or activity of p53 were investigated by co-immunoprecipitation, immunofluorescence, and immunoblot analyses. Levels of small nucleolar RNA H/ACA box 18-like 5 (SNORA18L5) were quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: We identified a low-frequency duplication at chromosome 15q13.3 strongly associated with risk of HBV-related HCC (overall P = 3.17 × 10-8; odds ratio, 12.02). Copy numbers of the 15q13.3 duplication correlated with the expression of SNORA18L5 in liver tissues. Overexpression of SNORA18L5 increased HCC cell proliferation and growth of xenograft tumors in mice; knockdown reduced HCC proliferation and tumor growth. SNORA18L5 overexpression in HepG2 and SMMC-7721 cells inhibited p53-dependent cell cycle arrest and apoptosis. Overexpression of SNORA18L5 led to hyperactive ribosome biogenesis, increasing levels of mature 18S and 28S ribosomal RNAs and causing the ribosomal proteins RPL5 and RPL11 to stay in the nucleolus, which kept them from binding to MDM2. This resulted in increased MDM2-mediated ubiquitination and degradation of p53. Levels of SNORA18L5 were increased in HCC tissues compared with nontumor liver tissues and associated with shorter survival times of patients. CONCLUSIONS: In a CNV-based GWAS, we associated duplication at 15q13.3 with increased risk of HBV-related HCC. We found SNORA18L5 at this location to promote HCC cell proliferation and tumor growth in mice. SNORA18L5 increases ribosome biogenesis, facilitates ribosomal RNA maturation, and alters localization of RPL5 and RPL11, allowing for increased MDM2-mediated proteolysis of p53 and cell cycle arrest.

Genome-Wide Association Study Identifies a New Locus at 7q21.13 Associated with Hepatitis B Virus-Related Hepatocellular Carcinoma.

Purpose: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. In China, chronic hepatitis B virus (HBV) infection remains the major risk factor for HCC. In this study, we performed a genome-wide association study (GWAS) among Chinese populations to identify novel genetic loci contributing to susceptibility to HBV-related HCC.Experimental Design: GWAS scan is performed in a collection of 205 HBV-related HCC trios (each trio includes an affected proband and his/her both parents), and 355 chronic HBV carriers with HCC (cases) and 360 chronic HBV carriers without HCC (controls), followed by two rounds of replication studies totally consisting of 3,796 cases and 2,544 controls.Results: We identified a novel association signal within the CDK14 gene at 7q21.13 (index rs10272859, OR = 1.28, P = 9.46 × 10-10). Furthermore, we observed that the at-risk rs10272859[G] allele was significantly associated with higher mRNA expression levels of CDK14 in liver tissues. Chromosome conformation capture assays in liver cells confirmed that a physical interaction exists between the promoter region of CDK14 and the risk-associated SNPs in strong linkage disequilibrium with the index rs10272859 at 7q21.13. This index rs10272859 also showed significant association with the survival of HCC patients.Conclusions: Our findings highlight a novel locus at 7q21.13 conferring both susceptibility and prognosis to HBV-related HCC, and suggest the CDK14 gene to be the functional target of the 7q21.13 locus. Clin Cancer Res; 24(4); 906-15. ©2017 AACR.