Identification of tumor antigens and immune subtypes of hepatocellular carcinoma for mRNA vaccine development

BACKGROUND mRNA vaccines have been investigated in multiple tumors,but limited studies have been conducted on their use for hepatocellular carcinoma(HCC).AIM To identify candidate mRNA vaccine antigens for HCC and suitable subpopu-lations for mRNA vaccination.METHODS Gene expression profiles and clinical information of HCC datasets were obtained from International Cancer Genome Consortium and The Cancer Genome Atlas.Genes with somatic mutations and copy number variations were identified by cBioPortal analysis.The differentially expressed genes with significant prognostic value were identified by Gene Expression Profiling Interactive Analysis 2 website analysis.The Tumor Immune Estimation Resource database was used to assess the correlation between candidate antigens and the abundance of antigen-presenting cells(APCs).Tumor-associated antigens were overexpressed in tumors and associated with prognosis,genomic alterations,and APC infiltration.A consensus cluster analysis was performed with the Consensus Cluster Plus package to identify the immune subtypes.The weighted gene coexpression network analysis(WGCNA)was used to determine the candidate biomarker molecules for appropriate populations for mRNA vaccines.immune subtypes showed distinct cellular and clinical characteristics.The IS1 and IS3 immune subtypes were immunologically“cold”.The IS2 and IS4 immune subtypes were immunologically“hot”,and the immune checkpoint genes and immunogenic cell death genes were upregulated in these subtypes.IS1-related modules were identified with the WGCNA algorithm.Ultimately,five hub genes(RBP4,KNG1,METTL7A,F12,and ABAT)were identified,and they might be potential biomarkers for mRNA vaccines.CONCLUSION AURKA,CCNB1,CDC25C,CDK1,TRIP13,PES1,MCM3,PPM1G,NEK2,KIF2C,PTTG1,KPNA2,and PRC1 have been identified as candidate HCC antigens for mRNA vaccine development.The IS1 and IS3 immune subtypes are suitable populations for mRNA vaccination.RBP4,KNG1,METTL7A,F12,and ABAT are potential biomarkers for mRNA vaccines.

Immunohistochemical detection of proliferating cell nuclear antigen in hepatocellular carcinoma

ＩｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｃａｌｄｅｔｅｃｔｉｏｎｏｆｐｒｏｌｉｆｅｒａｔｉｎｇｃｅｌｎｕｃｌｅａｒａｎｔｉｇｅｎｉｎｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａＷＡＮＧＤｏｎｇ１，ＳＨＩＪｉｎｇＱｕａｎ２ａｎｄＬＩＵＦｅｎｇＸｕａｎ３Ｓｕ...

Diagnostic value of cancer-testis antigen mRNA in peripheral blood from hepatocellular carcinoma patients

AIM:To evaluate the diagnostic value of cancer-testis antigen(CTA) mRNA in peripheral blood samples from hepatocellular carcinoma(HCC) patients.METHODS:Peripheral blood samples were taken from 90 patients with HCC before operation.Expression of melanoma antigen-1(MAGE-1),synovial sarcoma X breakpoint-1(SSX-1),and cancer-testis-associated protein of 11 kDa(CTp11) mRNA in peripheral blood mononuclear cells(PBMC) was tested by nested reverse transcriptspolymerase chain reaction(RT-PCR).Serum α-fetoprotein(AFP) in these patients was also determined.RESULTS:The positive rate of MAGE-1,SSX-1 and CTp11 transcripts was 37.7%,34.4%,31.1% in PBMC samples,and 74.4%,73.3%,62.2% in their resected tumor samples,respectively.The positive rate for at least one of the transcripts of three CTA genes was 66.7% in PBMC samples and 91.1% in their resected tumor samples.MAGE-1,SSX-1 and/or CTp11 mRNA were not detected in the PBMC of those patients from whom the resected tumor samples were MAGE-1,SSX-1 and/or CTp11 mRNA negative,nor in the PBMC samples from 20 healthy donors and 10 cirrhotic patients.Among the 90 patients,the serum AFP in 44 patients met the general diagnostic standard(AFP > 400 μg/L) for HCC,and was negative(AFP ≤ 20 μg/L) or positive with a low concentration(20 μg/L < AFP ≤ 400 μg/L) in the other patients.The positive rate for at least one of the transcripts of three CTA genes in PBMC samples from the AFP negative or positive patients with a low concentration was 69.2% and 45.0%,respectively.Of the 90 patients,71(78.9%) were diagnosed as HCC by nested RT-PCR and serum AFP.Although the positive rate for at least one of the transcripts of three CTA genes in PBMC samples from 53 patients at TNM stage or was obviously higher than that in PBMC samples from 37 patients at stage or(77.9% vs 51.4%,P = 0.010),the CTA mRNA was detected in 41.7% and 56.0% of PBMC samples from HCC patients at stages andrespectively.CONCLUSION:Detecting MAGE-1,SSX-1 and CTp11 mRNA in PBMC improves the total diagnostic rate of HCC.

The expression of c-kit and proliferating cell nuclear antigen in oval cells of rats with hepatocellular carcinoma

OBJECTIVE: To study the relationship between oval cells and primary hepatocarcinoma and the expression of c-kit and proliferating cell nuclear antigen (PCNA) in oval cells of rats with hepatocellular carcinoma. METHODS: A hundred and twenty clean SD rats were divided into three groups: normal group, cancer-induction group and intervention group. The normal group was fed with standard forage while the rest two groups were fed with 3'-methyl-2-methylamino-azobenzene (DAB) to induce carcinoma for 14 weeks and then fed with standard forage and water. Uscharidin was injected abdominally to the intervention group from the first week to the 14th week. All rats were killed and biopsy specimens were taken from the left and right liver lobes for immunohistochemical staining of c-kit and PCNA on the 2nd, 4th, 6th, 8th, 10th, 12th, 14th, 16th, 18th, 20th, 22nd, and 24th week. RESULTS: From the 2nd to 14th week after liver infection, c-kit positive cells, mainly oval cells were found in the portal area in the carcinoma-induction group and dotted positive pigmentations in liver lobules. In the 22nd week, a large number of cancerous nodes occurred and nuclei heteromorphi-m was apparent; the number of positive cell decreased but positive cells could be sparsely observed in cancerous nodes. In the 2nd week of the carcinoma-induction process, PCNA positive cells were oval cells in the portal area. In the 4th week, a lot of hepatic cells were positively stained, especially in the central vein area. In the 6th week, PCNA positive cells could be seen in the lobules of the liver. In the 8th week, the number of PCNA cells decreased comparatively. From the 10th to 14th week, oval cells in the portal area were still over-expressed. From the 16th to 24th week, a large number of cancerous nodes occurred and PCNA was over-expressed in some of them. In necrotic cancerous nodes, the para-cancerous PCNA positive cells were sparsely distributed and their number was less than that of PCNA positive cells of cancerous tissues. CONCLUSIONS: Hepatic stem cells originating from the terminal biliary plexus of the portal area are involved in the development of hepatocarcinoma because c-kit positive cells expressed in cancerous nodes, accompany the whole process of the development. In the middle inflammatory period of carcinoma-induction, the expression of PCNA in hepatic cells peaked, but the index decreased in the late inflammatory period and in the proliferated fibrosis stage. The expression of PCNA is a tortuous process, going up, down, then up again from normal tissues to cancerous tissues. Combined with pathological findings, PCNA can be considered as a warning index for carcinomatous cells.

Intrahepatic distribution of hepatitis B virus antigens in patients with and without hepatocellular carcinoma

AIM: To study the intrahepatic expression of hepatitis B surface antigen(HBs Ag) and hepatitis B core antigen(HBc Ag) in chronic hepatitis B patients with and without hepatocellular carcinoma. METHODS: A total of 33 chronic hepatitis B patients(mean age of 40.3 ± 2.5 years), comprising of 14 HBe Ag positive and 19 HBe Ag negative patients; and 13 patients with hepatitis B virus related hepatocellular carcinoma(mean age of 49.6 ± 4.7 years), were included in our study. Immunohistochemical staining for HBc Ag and HBs Ag was done using standard streptavidin-biotin-immunoperoxidase technique on paraffin-embedded liver biopsies. The HBc Agand HBs Ag staining distributions and patterns were described according to a modified classification system. RESULTS: Compared to the HBe Ag negative patients, the HBe Ag positive patients were younger, had higher mean HBV DNA and alanine transaminases levels. All the HBe Ag positive patients had intrahepatic HBc Ag staining; predominantly with "diffuse" distribution(79%) and "mixed cytoplasmic/nuclear " pattern(79%). In comparison, only 5% of the HBe Ag-negative patients had intrahepatic HBc Ag staining. However, the intrahepatic HBs Ag staining has wider distribution among the HBe Ag negative patients, namely; majority of the HBe Ag negative cases had "patchy" HBs Ag distribution compared to "rare" distribution among the HBe Ag positive cases. All but one patient with HCC were HBe Ag negative with either undetectable HBV DNA or very low level of viremia. Intrahepatic HBc Ag and HBs Ag were seen in 13(100%) and 10(77%) of the HCC patients respectively. Interestingly, among the 9 HCC patients on anti-viral therapy with suppressed HBV DNA, HBc Ag and HBs Ag were detected in tumor tissues but not the adjacent liver in 4(44%) and 1(11%) patient respectively. CONCLUSION: Isolated intrahepatic HBc Ag and HBs Ag can be present in tumors of patients with suppressed HBV DNA on antiviral therapy; that may predispose them to cancer development.

Expression of A, G and B melanoma antigen genes in human hepatocellular carcinoma

Objective: To observe the expression of the A melano- ma antigen (MAGE), G melanoma antigen (GAGE) and B melanoma antigen (BAGE) genes in human hepatocellular carcinoma cell lines. Methods: The MAGE-1, MAGE-3, GAGE1-8, GAGE1-2 and BAGE mRNA lever in hepatocellular carcinoma cell lines SMMC-7721, QQY-7701, BEL- 7402 were studied by reverse transcription polymer- ase chain reaction and were compared with biopsied liver tissues. Results: MAGE-1 and BAGE mRNA were expressed in SMMC-7721, MAGE-3 and BAGE in QGY-7701, MAGE-1 and GAGE1-2 in BEL-7402. None of these genes was expressed in biopsied liver tissues. Conclusions: MAGE-1, MAGE-3, GAGE1-8, GAGE1-2 and BAGE were expressed in hepatocellu- lar carcinoma cell lines, respectively. These tumor- specific antigens can be used as molecular markers and possible targets of immunotherapy for patients with hepatocellular carcinoma.

Prostate-specific membrane antigen expression in hepatocellular carcinoma,cholangiocarcinoma,and liver cirrhosis

BACKGROUND Primary liver cancer includes three subtypes:Hepatocellular carcinoma(HCC),intrahepatic cholangiocarcinoma(CCA),and combined hepatocellular carcinoma.Patients with primary liver cancer experienced poor prognosis and high mortality,so early detection of liver cancer and improved management of metastases are both key strategies to reduce the death toll from liver cancer.Prostate-specific membrane antigen(PSMA)expression in the tumor-associated neovasculature of nonprostate malignancies including liver cancer has been reported recently,but conclusive evidence of PSMA expression based on the pathological type of liver cancer remains limited.AIM To study the expression of PSMA in HCC,CCA,and liver cirrhosis.METHODS A total of 446 formalin-fixed paraffin-embedded(FFPE)liver tumor and liver cirrhosis tissue samples were obtained retrospectively from the Pathology Department of Tongji Hospital.Immunohistochemistry was used to detect PSMA expression in these 446 FFPE liver biopsy specimens(213 HCC,203 CCA,and 30 liver cirrhosis).The tumor compartment and the associated neovascular endothelium were separately analyzed.PSMA expression was examined by two certified pathologists,and the final results were presented in a 4-point scoring system(0-3 points).Correlation between PSMA expression and clinicopathological information was also assessed.RESULTS PSMA was expressed primarily in the neovascular endothelium associated with tumors.The positive rate of PSMA staining in HCC was significantly higher than that in CCA(86.8%vs 79.3%;P=0.001)but was only 6.6%in liver cirrhosis(P=0.000).HCC cases had more 3-score PSMA staining than CCA had(89/213,41.8%vs 35/203,17.2%;P=0.001).PSMA expression correlated positively with the stage and grade of HCC and CCA.In both liver cancer subtypes,there were more PSMA+cases in stages III–V diseases than in stages I and II.High staining intensity of PSMA was more frequently observed in liver cancers at high grade and advanced stage.There was no significant association of PSMA expression with sex,age,region,α-fetoprotein,hepatitis B surface antigen,or tumor size in both tumor subtypes.CONCLUSION Neovascular PSMA may be a promising marker to differentiate HCC from liver cirrhosis and a prognostic marker for anti-tumor angiogenesis therapy for HCC.

Advancement of chimeric antigen receptor-natural killer cells targeting hepatocellular carcinoma

With the advance of genome engineering technology,chimeric antigen receptors(CARs)-based immunotherapy has become an emerging therapeutic strategy for tumors.Although initially designed for T cells in tumor immunotherapy,CARs have been exploited to modify the function of natural killer(NK)cells against a variety of tumors,including hepatocellular carcinoma(HCC).CAR-NK cells have the potential to sufficiently kill tumor antigen-expressing HCC cells,independent of major histocompatibility complex matching or prior priming.In this review,we summarize the recent advances in genetic engineering of CAR-NK cells against HCC and discuss the current challenges and prospects of CAR-NK cells as a revolutionary cellular immunotherapy against HCC.

Autoantibodies against tumor-associated antigens for detection of hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is one of the most common tumors worldwide. The survival rate after the onset of symptoms is generally less than one year for the late presentation of HCC, and reliable tools for early diagnosis are lacking. Therefore, novel biomarkers for the early detection of HCC are urgently required. Recent studies show that the abnormal release of proteins by tumor cells can elicit humoral immune responses to self-antigens called tumor-associated antigens(TAAs). The corresponding autoantibodies can be detected before the clinical diagnosis of cancer. Therefore, there is growing interest in using serum autoantibodies as cancer biomarkers. In this review, we focus on the advances in research on autoantibodies against TAAs as serum biomarker for detection of HCC, the mechanism of the production of TAAs, and the association of autoantibodies with patients' clinical characteristics.

c-Met-targeted chimeric antigen receptor T cells inhibit hepatocellular carcinoma cells in vitro and in vivo

c-Met is a hepatocyte growth factor receptor overexpressed in many tumors such as hepatocellular carcinoma(HCC).Therefore,c-Met may serve as a promising target for HCC immunotherapy.Modifying T cells to express c-Met-specific chimeric antigen receptor(CAR)is an attractive strategy in treating c-Met-positive HCC.This study aimed to systematically evaluate the inhibitory effects of 2^(nd)-and 3^(rd)-generation c-Met CAR-T cells on hepatocellular carcinoma(HCC)cells.Here,2^(nd)-and 3^(rd)-generation c-Met CARs containing an anti-c-Met singlechain variable fragment(scFv)as well as the CD28 signaling domain and CD3ζ(c-Met-28-3ζ),the CD137 signaling domain and CD3ζ(c-Met-137-3ζ),or the CD28 and CD137 signaling domains and CD3ζ(c-Met-28-137-3ζ)were constructed,and their abilities to target c-Met-positive HCC cells were evaluated in vitro and in vivo.All c-Met CARs were stably expressed on T cell membrane,and c-Met CAR-T cells aggregated around c-Met-positive HCC cells and specifically killed them in vitro.c-Met-28-137-3ζCAR-T cells secreted more interferon-gamma(IFN-γ)and interleukin 2(IL-2)than c-Met-28-3ζCAR-T cells and c-Met-137-3ζCAR-T cells.Compared with c-Met low-expressed cells,c-Met CAR-T cells secreted more cytokines when co-cultured with c-Met high-expressed cells.Moreover,c-Met-28-137-3ζCAR-T cells eradicated HCC more effectively in xenograft tumor models compared with the control groups.This study suggests that 3^(rd)-generation c-Met CAR-T cells are more effective in inhibiting c-Met-positive HCC cells than 2^(nd)-generation c-Met CAR-T cells,thereby providing a promising therapeutic intervention for c-Met-positive HCC.

Clinical relevance of increased serum preneoplastic antigen in hepatitis C-related hepatocellular carcinoma

BACKGROUND The prognosis of hepatocellular carcinoma(HCC)patients remains poor despite advances in treatment modalities and diagnosis.It is important to identify useful markers for the early detection of HCC in patients.Preneoplastic antigen(PNA),originally reported in a rat carcinogenesis model,is increased in the tissues and serum of HCC patients.AIM To determine the diagnostic value of PNA for discriminating HCC and to characterize PNA-positive HCC.METHODS Patients with hepatitis C virus(HCV)-related hepatic disorders were prospectively enrolled in this study,which included patients with hepatitis,with cirrhosis,and with HCC.A novel enzyme-linked immunosorbent assay was developed to measure serum PNA concentrations in patients.RESULTS Serum PNA concentrations were measured in 89 controls and 141 patients with HCV infections(50 hepatitis,44 cirrhosis,and 47 HCC).Compared with control and non-HCC patients,PNA was increased in HCC.On receiver operating characteristic curve analysis,the sensitivity of PNA was similar to the HCC markers des-γ-carboxy-prothrombin(DCP)andα-fetoprotein(AFP),but the specificity of PNA was lower.There was no correlation between PNA and AFP and a significant but weak correlation between PNA and DCP in HCC patients.Importantly,the correlations with biochemical markers were completely different for PNA,AFP,and DCP;glutamyl transpeptidase was highly correlated with PNA,but not with AFP or DCP,and was significantly higher in PNA-high patients than in PNA-low patients with HCV-related HCC.CONCLUSION PNA may have the potential to diagnose a novel type of HCC in which glutamyl transpeptidase is positively expressed but AFP or DCP is weakly or negatively expressed.

Combination of squamous cell carcinoma antigen immunocomplex and alpha-fetoprotein in mid-and long-term prediction of hepatocellular carcinoma among cirrhotic patients

BACKGROUND The combination of alpha-fetoprotein(AFP)and squamous cell carcinoma antigen immunocomplex(SCCA-IgM)have been proposed for its use in the screening of hepatocellular carcinoma(HCC).Current screening programs for all cirrhotic patients are controversial and a personalized screening is an unmet need in the precision medicine era.AIM To determine the role of the combination of SCCA-IgM and AFP in predicting mid-and long-term appearance of HCC.METHODS Two-hundred and three cirrhotic patients(Child A 74.9%,B 21.2%,C 3.9%)were followed-up prospectively every six months to screen HCC by ultrasound and AFP according to European Association for the Study of the Liver guidelines.The estimation cohort was recruited in Italy(30.5%;62/203)and validation cohort from Spain(69.5%;141/203).Patients underwent to evaluate SCCA-IgM by enzyme-linked immunosorbent assay(Hepa-IC,Xeptagen,Italy)and AFP levels at baseline.Patients were followed-up for 60 mo,being censored at the time of the appearance of HCC.RESULTS There were 10.8%and 23.1%of HCC development at two-and five-years followup.Patients with HCC showed higher levels of SCCA-IgM than those without it(425.72±568.33 AU/mL vs 195.93±188.40 AU/mL,P=0.009)during the fiveyear follow-up.In multivariate analysis,after adjusting by age,sex,aspartate transaminase and Child-Pugh,the following factors were independently associated with HCC:SCCA-IgM[Hazard ratio(HR)=1.001,95%CI:1.000-1.002;P=0.003],AFP(HR=1.028,95%CI:1.009-1.046;P=0.003)and creatinine(HR=1.56495%CI:1.151-2.124;P=0.004).The log-rank test of the combination resulted in 7.488(P=0.024)in estimation cohort and 11.061(P=0.004)in the validation cohort,and a 100%of correctly classified rate identifying a low-risk group in both cohorts in the two-year follow-up.CONCLUSION We have constructed a predictive model based on the combination of SCCA-IgM and AFP that provides a new HCC screening method,which could be followed by tailored HCC surveillance for individual patients,especially for those cirrhotic patients belonging to the subgroup identified as low-risk of HCC development.

High-dose hepatitis B immunoglobulin therapy in hepatocellular carcinoma with hepatitis B virus-DNA/hepatitis B e antigen-positive patients after living donor liver transplantation

AIM: To investigate the impact of high-dose hepatitis B immunoglobulin(HBIG) on hepatocellular carcinoma(HCC) and hepatitis B virus(HBV) recurrence and overall survival after living donor liver transplantation(LDLT).METHODS: We investigated 168 patients who underwent LDLT due to HCC, and who were HBV-DNA/hepatitis B e antigen(HBe Ag)-positive, from January 2008 to December 2013. After assessing whether the patients met the Milan criteria, they were assigned to the low-dose HBIG group and high-dose HBIG group. Using the propensity score 1:1 matching method, 38 and 18 pairs were defined as adhering to and not adhering to the Milan criteria. For each pair, HCC recurrence, HBV recurrence and overall survival were analyzed by the Kaplan-Meier method and the log rank test according to the HBIG dose. RESULTS: Among those who met the Milan criteria, the 6-mo, 1-year, and 3-year HCC recurrence-free survival rates were 88.9%, 83.2%, and 83.2% in the low-dose HBIG group and 97.2%, 97.2%, and 97.2% in the high-dose HBIG group, respectively(P = 0.042).In contrast, among those who did not meet the Milan criteria, HCC recurrence did not differ according to the HBIG dose(P = 0.937). Moreover, HBV recurrence and overall survival did not differ according to the HBIG dose among those who met(P = 0.317 and 0.190, respectively) and did not meet(P = 0.350 and 0.987, respectively) the Milan criteria. CONCLUSION: High-dose HBIG therapy can reduce HCC recurrence in HBV-DNA/HBe Ag-positive patients after LDLT.

Neuron-glial antigen 2 overexpression in hepatocellular carcinoma predicts poor prognosis

AIM:To investigate whether neuron-glial antigen 2(NG2) could be an effective prognostic marker in hepatocellular carcinoma(HCC).METHODS:NG2 expression was semi-quantitatively scored from the immunohistochemistry(IHC) data based on the number of positive cells and the staining intensity.A total of 132 HCC specimens and 96 adjacent noncancerous tissue samples were analyzed by IHC for NG2 protein expression.To confirm the NG2 expression levels observed by IHC,we measured NG2 expression in 30 randomly selected tumor and adjacent noncancerous tissue samples by quantitative real-time polymerase chain reaction and Western blot.The correlations between NG2 protein expression and the clinicopathological features of HCC patients were analyzed using the χ2 test.To assess the prognostic value of NG2 for HCC,the association between NG2 expression and survival was analyzed using the KaplanMeier method with the log-rank test.To further evaluate the prognostic value of NG2 expression,a Cox multivariate proportional hazards regression analysis was performed with all the variables to derive risk estimates related to disease-free and overall survival and to control for confounders.RESULTS:High NG2 expression was observed in significantly more primary tumor samples(63.6%; 84/132) compared with the adjacent noncancerous tissue samples(28.1%; 27/96)(P < 0.0001).Moreover,high NG2 protein expression was closely associated with tumor differentiation(χ2 = 9.436,P = 0.0089),recurrence(χ2 = 5.769,P = 0.0163),tumor-nodemetastasis(TNM) stage(χ2 = 8.976,P = 0.0027),and invasion(χ2 = 5.476,P = 0.0193).However,no significant relationship was observed between NG2 protein expression in HCC and other parameters,such as age,sex,tumor size,serum alpha fetoprotein(AFP),tumor number,or tumor capsule.The log-rank test indicated a significant difference in the overall survival of HCC patients with high NG2 expression compared with those with low NG2 expression(29.2% vs 9.5%,P < 0.001).Moreover,NG2 expression in HCC tissue significantly correlated with disease-free survival(15.2% vs 6.7%,P < 0.001).Multivariate analysis showed that NG2 expression(HR = 2.035,P = 0.002),serum AFP(HR = 1.903,P = 0.003),TNM stage(HR = 2.039,P = 0.001),and portal vein invasion(HR = 1.938,P = 0.002) were independent prognostic indicators for OS in HCC patients.Furthermore,NG2 expression(HR = 1.974,P = 0.003),serum AFP(HR = 1.767,P = 0.008),TNM stage(HR = 2.078,P = 0.001),tumor capsule(HR = 0.652,P = 0.045),and portal vein invasion(HR = 1.941,P = 0.002) were independent prognostic indicators for DFS in HCC patients.CONCLUSION:The up-regulation of NG2 is associated with poor prognosis in HCC.Therefore,NG2 could be useful as an additional prognostic marker to increase the resolution of traditional approaches.

Significance of hepatitis B virus surface antigen, hepatitis C virus expression in hepatocellular carcinoma and pericarcinomatous tissues

AIM: To investigate the correlation between hepatitis B virus surface antigen (HBsAg), hepatitis C virus (HCV) expression in hepatocellular carcinoma (HCC), the HAI score of the noncancerous region of the liver and the serum Alpha fetoprotein (AFP) level.METHODS: The patterns of HBsAg and HCV in 100 cases of HCC and their surrounding liver tissues were studied on paraffin-embedded sections with immuno-histochemistry, the histological status was determined by one pathologist and one surgeon simultaneously using the hepatitis activity index (HAI) score, and AFP was detected by radioimmunity. The study included 100 consecutive patients who underwent curative resection for HCC. Based on HBsAg and HCV expression, the patients were classified into 4 groups: patients positive for HBsAg (HBsAg group), patients positive for HCV (HCV group), patients negative for both HCV and HBsAg (NBNC group) and patients positive for both HBsAg and HCV (BC group).RESULTS: The BC group had significantly higher HAI scores than the other three groups. (BC > HCV > HBsAg > NBNC). HBV and HCV virus infection was positively correlated with HAI (rs = 0.39, P = 0.0001). The positive rate of AFP (85.7%) and the value of AFP (541.2 ng/mL) in the group with HBV and HCV co-infection were the highest among the four groups. The positive rate (53.3%) of AFP and the value of AFP ( 53.3 ng/mL) in the group with none-infection of HBV and HCV were the lowest. HBV and HCV virus infection was positively correlated with AFP(rs = 0.38, P = 0.0001). CONCLUSION: The AFP increase in patients with liver cancer was positively correlated with the infection of HBV and HCV. The serum AFP elevation by the infection of HBV and HCV is one of mechanisms which lead to hepatocarcinogenesis, and the antivirus intervening treatment of hepatitis is significant for the prognosis of liver cancer. From our Spearman’s rank correlation analysis, we can conclude that the severity of virally induced inflammation is correlated with HBsAg and HCV expression in HCC tissues and noncancerous tissues. Prior co-infection of HBV in HCV patients may be an adverse risk factor for intrahepatic inflammation.

Expression of HLA-DR antigen in hepatocellular carcinoma and its up-regulation by interferon

Objective:To observetheexpressionof humanleukocyticantigenDR(HLA-DR)inprimaryhepatocellular carcinoma(HCC)anditsup-regulationby interferon(IFN).Methods:The expressionof HLA-DRin46specimensof humanHCCtissues,4humanHCCcelllines(SMMC-7721,HCC-9204,BEL-7402andHHCC)anda humanhepatocyte celllineQZGwas respectivelydetectedby immunohistochemicalABCstainingandflowcytometry.Theexpressionof HLA-DRinthe5celllineswasdetectedby ELISAbeforeandafterthecellsweretreatedwithIFN-γor IFN-α.Results:Eighteenoutof46HCCtissues(39.1%)expressedHLA-DR,whereasallthenormallivertissuesimmediatelyadjacentto HCCtissueswereHLA-DR-negative.No obviousHLA-DR-positivestainingwas foundin allthe5celllines.The expressionof HLA-DRwas up-regulatedin allthe5celllinesafterIFN-γor IFN-αtreatment.The up-regulationof HLA-DRinQZGcellswaslessobviousthanthatinHCCcelllines.Theeffectof IFN-γwasmoresignificantthanthatof IFN-α.Conclusion:HCCtissuescanexpressHLA-DRto someextent,butHCCcelllinesdo notexpressdetectable HLA-DR.IFNcanup-regulateHLA-DRexpressioninHCCcells.

Immunotherapy for advanced or recurrent hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is associated with high morbidity and mortality,and is prone to intra-and extrahepatic metastasis due to the anatomical and functional characteristics of the liver.Due to the complexity and high relapse rate associated with radical surgery or radiofrequency ablation,immune checkpoint inhibitors(ICIs)are increasingly being used to treat HCC.Several immunotherapeutic agents,along with their combinations,have been clinically approved to treat advanced or recurrent HCC.This review discusses the leading ICIs in practice and those currently undergoing randomized phase 1-3 trials as monotherapy or combination therapy.Furthermore,we summarize the rapidly developing alternative strategies such as chimeric antigen receptor-engineered T cell therapy and tumor vaccines.Combination therapy is a promising potential treatment option.These immunotherapies are also summarized in this review,which provides insights into the advantages,limitations,and novel angles for future research in establishing viable and alternative therapies against HCC.

肝细胞癌过继细胞疗法研究进展

肝细胞癌是严重威胁人类生存的一类疾病,中国肝细胞癌死亡率居世界第三。由于肝细胞癌发病早期无特异性症状,晚期的治疗方案又存在复发率高等的种种弊端,因此寻找行之有效的治疗新方法迫在眉睫。过继细胞疗法是一种细胞免疫疗法,在肝脏多样化和复杂的免疫微环境中,通过结合肿瘤特异性抗原或是非特异性的调节免疫平衡,过继细胞疗法在肝细胞癌的治疗中逐渐发挥出优势。在这篇综述中我们阐述了肝细胞癌靶向治疗常用生物标志物的形态结构及信号通路,探讨总结了近年来过继细胞疗法治疗肝细胞癌的研究手段及取得的临床效果,并对接下来的研究方向进行了展望。

CT23通过调节H6PD表达影响磷酸戊糖途径调控肝细胞癌细胞凋亡

目的:探讨癌-睾丸抗原23(CT23)参与磷酸戊糖途径(PPP)调控,促进肝细胞癌(HCC)细胞凋亡的分子机制。方法:通过全转录组测序、葡萄糖消耗检测、乳酸生成分析、还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)生成检测、活性氧(ROS)生成分析和线粒体示踪等方法探讨CT23与PPP的关系;蛋白质印记法(western blotting)及实时荧光定量PCR(RT-qPCR)检测敲低CT23的HCC细胞中已糖-6-磷酸脱氢酶(H6PD)表达量变化,TUNEL法分析细胞凋亡。结果:CT23与PPP有关;与control组相比,shCT23组HCC细胞葡萄糖消耗减少,乳酸生成水平降低,NADPH生成水平降低,ROS水平升高,细胞凋亡增加,H6PD mRNA水平降低,H6PD蛋白水平降低(均P<0.05);电镜下细胞形态发生变化并伴随线粒体损伤;与shCT23组相比,shCT23+H6PDOE组HCC细胞H6PD蛋白水平升高,葡萄糖消耗增多,乳酸生成水平升高,NADPH生成水平升高,细胞凋亡减少(均P<0.05)。结论:CT23通过H6PD增强PPP促进HCC细胞凋亡。

抗原提呈细胞亚群预测肝癌患者的免疫治疗疗效研究

背景原发性肝细胞癌(hepatocellular carcinoma,HCC)患者中免疫治疗应用广泛,但需寻找有效的疗效预测标志物,以此提供个体化和精准化的治疗。目的探讨抗原提呈细胞(antigen presenting cell,APC)亚群与HCC患者免疫治疗的疗效关系。方法前瞻性纳入2022年5月—2023年9月在解放军总医院第五医学中心初次接受免疫治疗的中晚期HCC患者,通过流式细胞仪检测免疫治疗前的B细胞/淋巴细胞(B/LYM)、巨噬细胞/所有核细胞(MA/TNC)和树突细胞/所有核细胞(DC/TNC)的比值。根据均值将患者分为高水平组和低水平组,并比较两组间无进展生存期(progression free survival,PFS),并通过Cox分析影响PFS的因素。最后利用ROC曲线预测疾病控制的效能。结果共纳入84例患者,中位年龄为57岁;男性58例,女性26例。APC亚群基线数据:B/LYM均值为16.54%±6.27%,MA/TNC均值为9.14%±2.87%,DC/TNC均值为0.051%±0.021%。Kaplan-Meier生存分析和Cox回归分析显示B/LYM高值组的mPFS显著高于低值组(5.90个月vs 5.50个月,P=0.019;HR=0.588,95%CI:0.358~0.966),MA/TNC低值组的mPFS显著高于高值组(7.20个月vs 5.40个月,P=0.034;HR=0.617,95%CI:0.374~1.019),DC/TNC高值组的mPFS显著高于低值组(6.30个月vs 5.50个月,P=0.026;HR=0.615,95%CI:0.378~0.955)。且高水平的MA/TNC是疾病进展的独立危险因素,高水平的DC/TNC、B/LYM是疾病进展的保护因素。基线B/LYM、MA/TNC、DC/TNC对HCC患者接受2周期治疗后的疾病控制情况而言,具有较好的预测效能,且联合应用时效能进一步提高,AUC分别为0.685、0.723、0.745、0.865。结论高DC/TNC、B/LYM水平和低MA/TNC水平的HCC患者,可能更容易从免疫治疗中获益。