Diagnostic value of 5 serum biomarkers for hepatocellular carcinoma with different epidemiological backgrounds:A large-scale,retrospective study

Objective:Hepatocellular carcinoma(HCC)is a lethal global disease that requires an accurate diagnosis.We assessed the potential of 5 serum biomarkers(AFP,AFU,GGT-II,GPC3,and HGF)in the diagnosis of HCC.Methods:In this retrospective study,we measured the serum levels of each biomarker using ELISAs in 921 participants,including 298 patients with HCC,154 patients with chronic hepatitis(CH),122 patients with liver cirrhosis(LC),and 347 healthy controls from 3 hospitals.Patients negative for hepatitis B surface antigen and hepatitis C antibody(called"NBNC-HCC")and patients positive for the above indices(called"HBV-HCC and HCV-HCC")were enrolled.The selected diagnostic model was constructed using a training cohort(n=468),and a validation cohort(n=453)was used to validate our results.Receiver operating characteristic analysis was used to evaluate the diagnostic accuracy.Results:Theα-L-fucosidase(AFU)/α-fetoprotein(AFP)combination was best able to distinguish NBNC-HCC[area under the curve:0.986(95%confidence interval:0.958–0.997),sensitivity:92.6%,specificity:98.9%]from healthy controls in the test cohort.For screening populations at risk of developing HCC(CH and LC),the AFP/AFU combination improved the diagnostic specificity for early-stage HCC[area under the curve:0.776(0.712–0.831),sensitivity:52.5%,specificity:91.6%in the test group].In all-stage HBV-HCC and HCV-HCC,AFU was also the best candidate biomarker combined with AFP[area under the curve:0.835(0.784–0.877),sensitivity 69.1%,specificity:87.4%in the test group].All results were verified in the validation group.Conclusions:The AFP/AFU combination could be used to identify NBNC-HCC from healthy controls and hepatitis-related HCC from at-risk patients.

Quantitative proteomic analysis for high-throughput screening of differential glycoproteins in hepatocellular carcinoma serum

Objective: Hepatocellular carcinoma(HCC) is a leading cause of cancer-related deaths. Novel serum biomarkers are required to increase the sensitivity and specificity of serum screening for early HCC diagnosis. This study employed a quantitative proteomic strategy to analyze the differential expression of serum glycoproteins between HCC and normal control serum samples.Methods: Lectin affinity chromatography(LAC) was used to enrich glycoproteins from the serum samples. Quantitative mass spectrometric analysis combined with stable isotope dimethyl labeling and 2D liquid chromatography(LC) separations were performed to examine the differential levels of the detected proteins between HCC and control serum samples. Western blot was used to analyze the differential expression levels of the three serum proteins.Results: A total of 2,280 protein groups were identified in the serum samples from HCC patients by using the 2D LC-MS/MS method. Up to 36 proteins were up-regulated in the HCC serum, whereas 19 proteins were down-regulated. Three differential glycoproteins, namely, fibrinogen gamma chain(FGG), FOS-like antigen 2(FOSL2), and α-1, 6-mannosylglycoprotein 6-β-N-acetylglucosaminyltransferase B(MGAT5B) were validated by Western blot. All these three proteins were up-regulated in the HCC serum samples.Conclusion: A quantitative glycoproteomic method was established and proven useful to determine potential novel biomarkers for HCC.

Identification of hub genes and their novel diagnostic and prognostic significance in pancreatic adenocarcinoma

Objective:The main reasons for the poor prognoses of pancreatic adenocarcinoma(PA)patients are rapid early-stage progression,advanced stage metastasis,and chemotherapy resistance.Identification of novel diagnostic and prognostic biomarkers of PA is therefore urgently needed.Methods:Three mRNA microarray datasets were obtained from the Gene Expression Omnibus database to select differentially expressed genes(DEGs).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses for hub genes were performed using DAVID.Correlations between expression levels of hub genes and cancer-infiltrating immune cells were investigated by TIMER.Cox proportional hazard regression analyses were also performed.Serum hub genes were screened using the HPA platform and verified for diagnostic value using ELISAs.Results:We identified 59 hub genes among 752 DEGs.GO analysis indicated that these 59 hub genes were mainly involved in the defense response to viruses and the type I interferon signaling pathway.We also discovered that RSAD2 and SMC4 were associated with immune cell infiltration in the PA microenvironment.Additionally,DLGAP5 mRNA might be used as an independent risk factor for the prognoses of PA patients.Furthermore,the protein encoded by ISG15,which exists in peripheral blood,was validated as a potential diagnostic biomarker that distinguished PA patients from healthy controls(area under the curve:0.902,95%confidence interval:0.819–0.961).Conclusions:Our study suggested that RSAD2 and SMC4 were associated with immune cell infiltration in the PA microenvironment,while DLGAP5 mRNA expression might be an independent risk factor for the survival prognoses of PA patients.Moreover,ELISAs indicated that serum ISG15 could be a potential novel diagnostic biomarker for PA.