Long non-coding RNA CASC7 is a promising serum biomarker for hepatocellular carcinoma.

BACKGROUND: At present, a large number of studies have found that long non-coding RNAs (lncRNAs) can be used as biomarkers for diagnosis and monitoring prognosis of hepatocellular carcinoma (HCC). The expression of lncRNA cancer susceptibility candidate 7 (CASC7) in HCC has rarely been studied. The purpose of this study was to explore the expression of CASC7 and its correlation with clinical features, and to further analyze its diagnostic value in HCC. METHODS: Serum samples were collected from 80 patients with HCC, 80 patients with chronic hepatitis B (CHB), and 80 healthy people. The expression level of serum CASC7 was detected by droplet digital PCR. Appropriate parametric and nonparametric tests were used for data analysis. RESULTS: The results showed that the expression of CASC7 in serum of patients with HCC was significantly higher than that of patients with CHB (median: 8.8 versus 2.2 copies/µl, p < 0.001) and healthy controls (median: 8.8 versus 3.8 copies/µl, p < 0.001). High expression of serum CASC7 was significantly correlated with tumor number (p = 0.005), intrahepatic metastasis (IM) (p < 0.001), tumor size (p = 0.007) and tumor-node-metastasis (TNM) stage (p = 0.008). The area under the curve (AUC) of CASC7 to distinguish HCC patients from CHB patients and healthy controls was 0.808 (95% CI: 0.742-0.874) at the cut-off value of 7.24 copies/µl with 63.8% sensitivity and 95.2% specificity. CONCLUSIONS: This study suggested that CASC7 was significantly up-regulated in serum of patients with HCC and closely related to tumor number, IM, tumor size and TNM stage, which may serve as a promising diagnostic biomarker.

A feedback loop between NONHSAT024276 and PTBP1 inhibits tumor progression and glycolysis in HCC by increasing the PKM1/PKM2 ratio.

Hepatocellular carcinoma (HCC) is one of the most common malignancies with a hallmark of aberrant metabolism. The mechanism of long noncoding RNAs (lncRNAs) underlying the aggressive behaviors and glycolysis of HCC is poorly understood. In this study, we identified, via microarray, novel lncRNA NONHSAT024276 as a potential tumor suppressor in HCC. The downregulation of NONHSAT024276 closely correlated with larger tumor volume and higher aspartate transaminase levels. Functional experiments were performed to verify the role of NONHSAT024276 in HCC progression, and the negative effects of NONHSAT024276 expression on cell proliferation and migration were identified. Mechanistically, NONHSAT024276 directly bound to polypyrimidine tract-binding protein 1 (PTBP1), downregulating it and forming a feedback loop. Furthermore, NONHSAT024276 increased the ratio of M1 and M2 isoforms of pyruvate kinase (PKM1/PKM2) and also obstructed the PTBP1/PKM-mediated glycolysis. Finally, the rescue assays confirmed that NONHSAT024276 functioned in HCC via downregulating PTBP1 to increase the PKM1/PKM2 ratio. Hence, this study supported a model in which NONHSAT024276 downregulated PTBP1 and formed a feedback loop to increase the PKM1/PKM2 ratio to inhibit glycolysis and progression of HCC, opening new prospects for preventing or treating HCC.

Screening and Functional Prediction of Key Candidate Genes in Hepatitis B Virus-Associated Hepatocellular Carcinoma.

BACKGROUND: The molecular mechanism by which hepatitis B virus (HBV) induces hepatocellular carcinoma (HCC) is still unknown. The genomic expression profile and bioinformatics methods were used to investigate the potential pathogenesis and therapeutic targets for HBV-associated HCC (HBV-HCC). METHODS: The microarray dataset GSE55092 was downloaded from the Gene Expression Omnibus (GEO) database. The data was analyzed by the bioinformatics software to find differentially expressed genes (DEGs). Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, ingenuity pathway analysis (IPA), and protein-protein interaction (PPI) network analysis were then performed on DEGs. The hub genes were identified using Centiscape2.2 and Molecular Complex Detection (MCODE) in the Cytoscape software (Cytoscape_v3.7.2). The survival data of these hub genes was downloaded from the Gene Expression Profiling Interactive Analysis (GEPIA). RESULTS: A total of 2264 mRNA transcripts were differentially expressed, including 764 upregulated and 1500 downregulated in tumor tissues. GO analysis revealed that these DEGs were related to the small-molecule metabolic process, xenobiotic metabolic process, and cellular nitrogen compound metabolic process. KEGG pathway analysis revealed that metabolic pathways, complement and coagulation cascades, and chemical carcinogenesis were involved. Diseases and biofunctions showed that DEGs were mainly associated with the following diseases or biological function abnormalities: cancer, organismal injury and abnormalities, gastrointestinal disease, and hepatic system disease. The top 10 upstream regulators were predicted to be activated or inhibited by Z-score and identified 25 networks. The 10 genes with the highest degree of connectivity were defined as the hub genes. Cox regression revealed that all the 10 genes (CDC20, BUB1B, KIF11, TTK, EZH2, ZWINT, NDC80, TPX2, MELK, and KIF20A) were related to the overall survival. CONCLUSION: Our study provided a registry of genes that play important roles in regulating the development of HBV-HCC, assisting us in understanding the molecular mechanisms that underlie the carcinogenesis and progression of HCC.

Effects of Blood Storage on Cell Population Data.

BACKGROUND: Cell population data (CPD) are leukocyte morphologic parameters, measured by automated hematology analyzers with VCS technology. Many studies have demonstrated that these parameters may have clinical utility for diagnosing or screening certain pathological conditions. This study is to investigate the effects of peripheral blood stored at room temperature on the CPD values and provide useful information about storage-induced CPD changes. METHODS: Venous blood samples from healthy donors kept at room temperature (18 - 25°C) for different time intervals were analyzed using a Coulter DxH800 hematology analyzer. The CPD data collected included mean cellular volume, conductivity and multiple angles of light scatters as well as their corresponding standard deviation for neutrophils, lymphocytes, and monocytes. Peripheral blood smears at each time interval were also prepared and examined microscopically. RESULTS: Peripheral blood kept at room temperature over time significantly affects the CPD values for neutrophils, lymphocytes, and monocytes. These CPD changes are correlated with the morphologic alterations observed under light microcopy, but detected much earlier. Some changes imitate clinical pathological conditions. For example, aged neutrophils showed decreased median angle light scatters, suggesting cytoplasmic degranulation, which can be seen in the case of myelodysplastic syndrome. CONCLUSIONS: This study provides valuable information about storage-induced CPD changes that can affect potential clinical application and interpretation of these automated digital morphologic parameters.

A Novel lncRNA NONHSAT053785 Acts as an Independent Risk Factor for Intrahepatic Metastasis of Hepatocellular Carcinoma.

PURPOSE: Long noncoding RNAs (lncRNAs) in body fluids have been considered as promising novel biomarkers for tumor-related diseases. The present study aimed to investigate the expression level of lncRNA NONHSAT053785 in serum and its correlation with clinical characteristics of hepatocellular carcinoma (HCC) patients. METHODS: The droplet digital PCR (ddPCR) was used to measure the serum levels of NONHSAT053785 in 112 HCC patients, 96 chronic hepatitis B (CHB) patients, and 99 healthy controls (HC). The correlation between NONHSAT053785 and clinical characteristics was analyzed by chi-square test and Spearman correlation test. The risk factors of intrahepatic metastasis (IM) were detected by univariate and multivariate analyses. Furthermore, the diagnostic value of NONHSAT053785 in HCC and its predictive ability in IM were evaluated by the receiver operating characteristic (ROC) curves. RESULTS: The level of NONHSAT053785 was significantly increased in the serum of HCC patients and was higher in HCC patients with IM as compared to those without. Additionally, the expression level of NONHSAT053785 was significantly related to IM, Child-Pugh classification, and peripheral blood indicators such as liver metabolic enzymes and positively correlated to IM, Barcelona Clinic Liver Cancer (BCLC) staging, and some peripheral blood indicators. Furthermore, the serum NONHSAT053785 was indicated as an independent predictor for IM in the elderly, non-smoking, drinking, and tumor size ≥5 cm subjects. The area under the ROC curve (AUC) was 0.801 (P <0.0001) for diagnosis of HCC and 0.678 (P =0.0015) for predicting IM. CONCLUSION: The increase in serum NONHSAT053785 levels was related to an increased risk of IM, and hence, may serve as a novel biomarker for the diagnosis of HCC and the prediction of IM.

Identification Of Novel lncRNAs For Detection Of HBV-Associated Hepatocellular Carcinoma.

PURPOSE: This study was conducted to investigate the differentially expressed profiles of long non-coding RNAs (lncRNAs) in HBV-associated HCC (HBV-HCC), which may serve as potential diagnostic biomarkers and therapeutic targets. METHODS: To examine the differentially expressed profiles of lncRNAs and mRNAs using microarray analysis, we collected 15 specimens: five HBV-associated HCC tissues, five paired adjacent peritumoral liver tissues (APLT), and five distant peritumoral liver tissues (DPLT). Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the biological roles and potential signaling pathways of these dysregulated mRNAs. In addition, lncRNA-mRNA co-expression network and signal transduction pathway network (Signal-net) were employed to further explore the potential target genes and roles of dysregulated lncRNAs in HBV-HCC pathogenesis. Finally, quantitative real-time PCR (qRT-PCR) was used to confirm the expression of six selected dysregulated lncRNAs. RESULTS: A total number of 719 lncRNAs and 3438 mRNAs were significantly more dysregulated in HBV-HCC tissues than in APLT and DPLT (fold change > 2, P < 0.05, FDR < 0.05). Additionally, 337 GO terms and 53 KEGG pathways were established to be significantly enriched. These dysregulated mRNAs were mainly enriched in metabolism-related biological processes. Additionally, lncRNA-mRNA coexpression network analysis showed that NONHSAT053785 is at the core of the network. Furthermore, the Signal-net analysis showed that CYP3A4 was gene with the highest degree. Finally, the data of five of the six selected differentially expressed lncRNAs were in agreement with the microarray data obtained by qRT-PCR verification. CONCLUSION: Our study revealed the differentially expressed profiles of lncRNAs and mRNAs for HBV-HCC, and five novel dysregulated lncRNAs were identified in HBV-HCC tissues. The aforementioned dysregulated lncRNAs may represent potential diagnostic biomarkers and therapeutic targets of HBV-HCC, which needs to be validated in future studies.

Evaluation of the Diagnostic Efficacy of Monocyte Parameters and MCP-1 to Distinguishing Active Tuberculosis from Latent Tuberculosis.

BACKGROUND: It is important to distinguish those with active Mycobacterium tuberculosis infection (ATB) from latent tuberculosis infection (LTBI) for monitoring and treating the disease. Monocytes play an important role and may undergo morphological changes against active TB infection. The aim of this study is to investigate the clinical usefulness of the monocyte morphometric parameters and monocyte chemoattractant protein-1 (MCP-1) to distinguish active tuberculosis (ATB) from latent tuberculosis infection (LTBI) and healthy controls (HC). METHODS: Peripheral blood was collected from 97 ATB patients, 113 LTBI patients, and 101 healthy controls. The monocyte morphometric parameters were obtained using a UniCel Coulter DxH800 system. MCP-1 level was determined by enzyme-linked immunosorbent assay method. Cutoff values were established based on receiver operator characteristic (ROC) curve analysis. RESULTS: Mean monocyte volume with its standard deviation, mean monocyte conductivity, and MCP-1 were significantly increased in ATB compared with LTBI and HC. ROC curve analyses showed that simultaneous measurements of mean monocyte volume with its standard deviation, mean monocyte conductivity and MCP-1 achieved good sensitivity and specificity (93.8% and 93.1%), which may be clinically useful. CONCLUSIONS: The findings using monocyte morphometric parameters and MCP-1 to distinguish ATB from LTBI with high sensitivity and specificity may be a potential parameter for clinical.

Circulating microRNA-122a as a diagnostic marker for hepatocellular carcinoma.

BACKGROUND: The purpose of this study was to evaluate the potential value of circulating miRNA-122a and miRNA-221 in the diagnosis of hepatocellular carcinoma. METHODS: Serum samples were obtained from 85 patients with hepatocellular carcinoma and 85 age-matched and sex-matched healthy volunteers. miRNAs were isolated from the serum samples, and alfa-fetoprotein levels were determined. Expression of miRNA-122a and miRNA-221 in cases and controls was quantified using U6 sn RNA as the internal control. The diagnostic value of miRNA-122a, miRNA-221, and alfa-fetoprotein was compared by receiver operating characteristic analysis. RESULTS: The serum miRNA-122a level in patients with hepatocellular carcinoma was significantly reduced in comparison with healthy controls and correlated with known risk factors for hepatocellular carcinoma. Circulating miRNA-221 in patients with hepatocellular carcinoma was higher compared with the control group, but the difference was not statistically significant. Receiver operating characteristic analysis revealed that the diagnostic power of miRNA-122a was suboptimal compared with serum alfa-fetoprotein. Further, the serum alfa-fetoprotein and miRNA-122a combined classifier resulted in performance similar to that of alfa-fetoprotein alone. CONCLUSION: The serum miRNA-122a level correlates with risk factors for hepatocellular carcinoma. However, use of miRNA-122a as a diagnostic tool for hepatocellular carcinoma is not superior to alfa-fetoprotein. Further analysis is needed to evaluate the diagnostic power of plasma miRNA-122a for hepatocellular carcinoma.

Clinical utility of a combination of lipoarabinomannan, 38-kDa, and 16-kDa antigens as a diagnosis tool for tuberculosis.

The aim of this study was to evaluate the diagnostic value of tests detecting antibodies against lipoarabinomannan (LAM), 38-kDa, and 16-kDa antigens for Mycobacterium tuberculosis (MTB). Sera from 160 tuberculosis (TB) patients and 150 non-TB healthy controls were subjected to simultaneous detection of antibodies against LAM, 38-kDa, and 16-kDa antigens using protein chips. The diagnostic value of the 3 TB antigens, alone or combined, was evaluated. Results showed that LAM and 38-kDa antigens had the highest positive rates in the TB patients. Tests showing any single positive antibody, 2 positive antibodies, and 3 positive antibodies had a sensitivity of 93.1%, 51.3%, and 15.6%, and a specificity of 81.3%, 96.6%, and 99.3%, respectively. The positive predictive value of tests showing any 2 positive antibodies and 3 positive antibodies was 94.2% and 96.1%, respectively. Combined detection of a selected panel of TB antibodies can improve the positive rates for TB diagnosis and can serve as an important aid to the diagnosis of TB especially extrapulmonary TB.

Association of DNA repair gene XRCC1 and XPD polymorphisms with genetic susceptibility to gastric cancer in a Chinese population.

BACKGROUND: DNA repair gene polymorphisms can contribute to susceptibility of human cancer, including gastric cancer. Three single nucleotide polymorphisms (SNPs) of xeroderma pigmentosum group D (XPD) and X-ray repair cross complement 1 (XRCC1) genes were genotyped in gastric cancer and control subjects in a population from Southwestern China for their association with susceptibility of gastric cancer risk. METHODS: 190 hospital-based cases and 180 matched controls were recruited and blood samples were collected from each of them and amplified with a PCR and DNA sequenced for XPD Asp312Asn, XRCC1 Arg194Trp, and XRCC1 Arg280Gln genotyping. RESULTS: Allelic association analysis of these three SNPs showed that the frequency of XRCC1 194Trp in gastric cancer case and the control was 17.2% and 7.3%, respectively, which was significantly associated with gastric cancer risk (OR=2.72, 95% CI: 1.04-7.24, p=0.027). Furthermore, XRCC1 194Trp allele increased gastric carcinoma risk in male patients with older age and distant metastasis of gastric cancer. In addition, XRCC1 Trp allele but not XRCC1 Arg allele was closely associated to development of gastric cardia carcinoma. However, other SNPs did not show an association with gastric cancer risk or other clinicopathologic data of the patients. CONCLUSION: XRCC1 194Trp allele significantly increased the risk of gastric cancer and also associated with risk of gastric cardia carcinoma and promoted distant metastasis of gastric cancer. Future study will verify these findings for use of this SNP as biomarker in gastric cancer.