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Liver cancer is the second leading cause of cancer-related deaths worldwide, with hepatocellular carcinoma (HCC) being the most common form of primary liver cancer. Glypican-3 (GPC3) is a cell membrane proteoglycan which is rarely expressed in normal adult tissues but is specifically upregulated in HCC, which makes GPC3 a reliable target for the diagnosis and treatment of HCC. The role of GPC3 in the regulation of cancer development through Wnt, YAP, hedgehog and other signaling pathways were reviewed in this article. GPC3-targeted therapies, such as monoclonal antibodies, bispecific antibodies, tumor vaccines, immunotoxins, CAR-T cells, and photosensitizer therapy were also summarized. These treatment methods offered promising approaches for HCC treatment and future treatment strategies for HCC based on GPC3 were prospected in this paper.
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Objective:To investigate the effect of plasma Glypican-4 level on the prognosis of children with pneumonia and acute respiratory distress syndrome(ARDS) and its predictive value.Methods:From April 2018 to April 2021, 92 children with pneumonia and ARDS admitted to the PICU of Baoding First Hospital for the first time were selected as the pneumonia+ ARDS group, 87 children with pneumonia admitted to the Pediatrics Department of this hospital were selected as the pneumonia group, and 95 children for physical examination in the same hospital were selected as control group.The pediatric critical illness score(PCIS)was used to score the children in pneumonia+ ARDS group and pneumonia group.The enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of Glypican-4 in plasma of three groups.Receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of plasma Glypican-4 in pneumonia complicated with ARDS, and the prognosis-related factors was analyzed by the Cox risk regression model.Results:The heart rate, pneumonia history, 28-day mortality rate, and plasma Glypican-4 in the pneumonia+ ARDS group were significantly higher than those in control group and pneumonia group, and PCIS score in pneumonia+ ARDS group was lower than that in pneumonia group ( P<0.05). According to the prognosis of pneumonia+ ARDS group on 28 days, the children were divided into survival group (74 cases) and death group (18 cases). Compared with the survival group[(3.92±0.31)μg/L], the plasma Glypican-4 level in death group[(4.78±0.35)μg/L] increased, whose difference was statistically significant( t=10.292, P<0.001). ROC curve analysis results showed that the area under the ROC curve of Glypican-4 predicting the prognosis of children with pneumonia combined with ARDS was 0.92, whose sensitivity was 94.2% and specificity was 88.3%, as well as the cut-off value was 4.11 μg/L.Multivariate Cox analysis showed that Glypican-4 level(high expression), heart rate (high frequency) and the history of pneumonia were prognostic risk factors affecting the prognosis of children with pneumonia and ARDS( P<0.05). Conclusion:The level of plasma Glypican-4 in children with pneumonia and ARDS significantly increased, and it is closely related to the prognosis of children with pneumonia and ARDS, and has high predictive value for the prognosis of children with pneumonia and ARDS.
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Objective:To study the glypican 3 (GPC3) fluorescent probe imagings in hepatocellular carcinoma (HCC) tissues and to determine its prognostic value in HCC patients.Methods:The data of 87 patients who were treated at the Affiliated Hospital of Southwest Medical University from January 2019 to August 2020 were retrospectively analyzed. There were 75 males and 12 females, with the age of (56.1±11.9) years. The expressions of GPC3 were measured by immunohistochemistry and by the fluorescent probe. The results obtained by these two tests were compared. Patients were followed up for recurrence after hepatectomy. Univariate and multivariate Cox regression analyses were used to analyze factors influencing recurrence-free survival.Results:Detection of the GPC3 expression by GPC3 fluorescence probe was consistent with the results obtained by immunohistochemical studies ( Kappa=0.84, P<0.001). The positive rates of GPC3 fluorescent probe was 79.3%(69/87), compared with 80.4%(70/87) by immunohistochemistry studies, with no significant difference between the two groups ( P>0.05). The patients were then divided into the low differentiation group ( n=30) and the middle high differentiation group ( n=57) by the degrees of tumor differentiation. The fluorescence intensity in the low differentiation group was 134.4(128.0, 144.7) a. u. which was significantly different from the middle high differentiation group of 84.8(0, 108.5)a.u. ( Z=-7.52, P<0.001). The median fluorescence intensity of 87 patients with HCC was 108.6 a. u.. Multivariate Cox regression analysis showed that patients with a GPC3 fluorescence intensity ≥108.6 a. u. ( HR=2.07, 95% CI: 1.21-3.53, P=0.008) had a significant increased risk of recurrence after hepatectomy. Conclusion:The expressions of GPC3 in HCC were consistent between the studies by using either a GPC3 specific fluorescent probe or immunohistochemistry studies. A GPC3 fluorescence intensity ≥108.6 a. u. was a risk factor of recurrence after hepatectomy in patients with HCC.
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Objective:To investigate the predictive value of a nomogram based on clinical factors and gadobenate dimeglumine (Gd-BOPTA)-enhanced MRI for predicting the expression of Glypican-3 (GPC-3) in hepatocellular carcinoma (HCC).Methods:The clinical and imaging data of 85 patients with HCC confirmed by pathology in the Provincial Hospital of Shandong First Medical University from July 2018 to June 2021 were retrospectively collected. All the patients underwent Gd-BOPTA-enhanced MRI scan before operation. According to the expression of GPC-3 by immunohistochemistry, the patients were divided into GPC-3 positive group (55 cases) and GPC-3 negative group (30 cases). The clinical data of patients were collected, including gender, age, hepatitis, cirrhosis, alpha-fetoprotein (AFP), alanine aminotransferase, aspartate aminotransferase, and glutamine transferase levels. The MRI qualitative signs including tumor margin, ring enhancement, intratumoral hemorrhage, enhanced capsule, and satellite nodules were reviewed. MRI quantitative parameters including the largest tumor diameter, Gd-BOPTA-enhanced tumor-to-liver parenchyma signal ratio (TLR) and tumor enhancement ratio (TER) in arterial phase (AP), portal venous phase (PP), and hepatobiliary phase (HBP) were calculated. The independent sample t-test or Mann-Whitney U test were used to compare the quantitative data between the two groups, and the χ2 test was used to compare the qualitative data between the two groups. Multivariate logistic regression analysis was used to identify the independent predictors of GPC-3 expression, and a nomogram model was established. The receiver operating characteristic (ROC) curves were used to evaluate the predictive performance of each independent factor and nomogram, and DeLong test was used to compare differences in area under the curve (AUC). Results:There were significant differences in AFP, tumor margin, intratumoral hemorrhage, and TLR-AP, TLR-PP and TLR-HBP between GPC-3 positive and negative groups (all P<0.05). Multivariate logistic regression results showed that AFP≥20 μg/L, intratumoral hemorrhage and TLR-HBP were independent predictors of GPC-3 positive expression in HCC (OR=3.816, 4.788, 0.001, all P<0.05). The preoperative clinical and Gd-BOPTA-enhanced MRI nomogram model for predicting GPC-3 expression in hepatocellular carcinoma was established. The AUC of AFP≥20 μg/L, intratumoral hemorrhage, TLR-HBP and nomogram model in predicting GPC-3 positive expression were 0.688, 0.697, 0.808, and 0.879, respectively. The AUC of nomogram model was significantly better than those of the other three single indicator ( Z=3.82, 4.13, 2.04, P<0.001,<0.001,=0.042). Conclusion:The nomogram model based on indicators of clinical and qualitative and quantitative Gd-BOPTA-enhanced MRI has better performance in predicting the expression of HCC GPC-3 before surgery, which is higher than those of each single indicator.
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Objective:To explore the expression of miR-6883-5p in the serum of bladder cancer patients and its effect on the migration and proliferation ability of bladder cancer cells.Methods:The real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression level of miR-6883-5p in the serum of 39 patients with bladder cancer and 39 healthy subjects, as well as the expression level of miR-6883-5p in normal bladder mucosal epithelial cells and bladder cancer cells. The bladder cancer cells with the lowest expression of miR-6883-5p were transfected with miR-6883-5p mimics or negative control (NC), namely miR-6883-5p group and NC group. Transwell migration experiment and cell counting kit-8 (CCK-8) colorimetric method were used to detect the migration and proliferation ability of cells transfected with miR-6883-5p. Bioinformatics software and dual luciferase reporter gene were used to predict and test the target genes of miR-6883-5p. qRT-PCR and Western blot were used to detect the expression levels of target genes in cells transfected with miR-6883-5p.Results:The expression level of miR-6883-5p in the serum of bladder cancer patients was significantly lower than that of healthy persons ( P<0.01). The expression level of miR-6883-5p in bladder cancer cells was significantly lower than that in normal bladder mucosal epithelial cells (all P<0.05), and the lowest expression of miR-6883-5p was in 5637 cells ( P<0.01). After transfection with miR-6883-5p, the migration and proliferation of cells in miR-6883-5p group were significantly lower than those in NC group (all P<0.05). Phosphatidyl alcohol proteoglycan-6 (GPC6) was the target gene of miR-6883-5p. After transfection with miR-6883-5p, the expression levels of GPC6 mRNA and protein in miR-6883-5p group significantly decreased ( P<0.01). Conclusions:miR-6883-5p is significantly low expressed in the serum of bladder cancer patients, and miR-6883-5p is related to the occurrence and development of bladder cancer; overexpression of miR-6883-5p can inhibit the migration and proliferation of bladder cancer cells; miR-6883-5p can inhibit the occurrence and development of bladder cancer by down-regulating the expression of GPC6.
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ABSTRACT Objective: Galanin is a neuropeptide which has effects not only on metabolic syndrome but also on reproduction. Glypican-4 is an adipokine associated with insulin sensitivity by interacting directly with the insulin receptor. This study evaluated serum concentrations of galanin and glypican-4 in relation with the hormonal profile as well as metabolic and cardiovascular risk factors in patients with and without polycystic ovary syndrome (PCOS). Subjects and methods: A total of 44 women with PCOS and 44 age-matched controls were eligible. Hirsutism scores, hormonal profile, metabolic and cardiovascular risk factors as well as galanin and glypican-4 levels were evaluated in each subject. Results: Women with PCOS exhibited lower levels of galanin (20.2 pg/mL versus 26.4 pg/mL, p = 0.002) and higher concentrations of glypican-4 (3.1 ng/mL versus 2.6 ng/mL, p < 0.001) than controls. Both adipokines were correlated positively with body mass index (BMI), insulin, triglyceride and Homeostasis Model Assessment (HOMA) index; glypican-4 also showed positive correlations with fasting blood glucose, free testosterone, modified Ferriman-Gallwey scores (p < 0.05). Multiple Linear Regression analyses showed that PCOS and BMI were the best predictors affecting galanin levels with a decreasing and increasing effect respectively; however BMI was the best predictor affecting glypican-4 levels with an increasing effect (p < 0.001). Conclusion: Galanin levels were lower and glypican-4 levels were higher in women with PCOS than controls. Further studies are needed to determine whether these adipokines could be used as additional markers for insulin sensitivity and lipid profile and whether they might play a role in the pathogenesis of PCOS, in which metabolic cardiovascular risks are increased.
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Humans , Female , Polycystic Ovary Syndrome/complications , Insulin Resistance , Galanin/blood , Glypicans/blood , Heart Disease Risk Factors , Cardiovascular Diseases/etiology , Body Mass Index , Case-Control Studies , Risk FactorsABSTRACT
ObjectiveTo investigate the effect of AU-rich element RNA-binding factor 1 (AUF1) on glypican 3 (GPC3) in hepatocellular carcinoma (HCC) and its possible mechanism. MethodsTCGA-HCC gene expression data were downloaded from Broad Institute Genome Data Analysis Center, and finally 371 HCC tissue samples with different etiologies and 50 adjacent tissue samples were included; LCI-HCC gene expression data were downloaded from GSE14520, and 214 patients with hepatitis B-associated HCC who had follow-up data were enrolled. A total of 35 primary liver cancer samples and corresponding adjacent tissue samples were collected from HCC patients who underwent radical surgery in Henan Provincial Cancer Hospital from 2009 to 2013. Immunohistochemistry was used to measure the protein expression of GPC3 and AUF1 in HCC tissue; Western Blot and qRT-PCR were used to measure the expression of GPC3 after AUF1 knockdown or overexpression in hepatoma cell lines; RNA-binding protein immunoprecipitation and RNA turnover assay were used to investigate the potential mechanism of AUF1 in regulating the expression of GPC3. The t-test was used for comparison of quantitative data between two groups, and the chi-square test was used for comparison of rates between two groups; the Kaplan-Meier method was used for survival analysis after surgery, and the log-rank test was used for comparison of survival rates. ResultsIn TCGA and LCI databases, the expression of GPC3 in HCC tissue was significantly higher than that in adjacent tissue (P＜0.05), and in TCGA database, the high expression of GPC3 was associated with the poor prognosis of HCC patients (P＜0.05). Immunohistochemistry showed that both GPC3 and AUF1 proteins are highly expressed in HCC tissue, with a positive expression rate of 77.1% (27/35) and 74.3% (26/35), respectively. In vitro experiment showed that AUF1 knockdown significantly reduced the expression of GPC3 in HepG2 and Huh-7 cells (P＜0.05), while AUF1 overexpression significantly increased the expression of GPC3 (P＜0.05). AUF1 protein could bind to GPC3 mRNA, and AUF1 knockdown reduced the stability of GPC3 mRNA. ConclusionAUF1 is an important post-transcriptional regulator of the GPC3 gene, and the abnormal high expression of AUF1 and GPC3 may be involved in the development and progression of HCC.
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Background and Aims@#Hepatocellular carcinoma (HCC) is a common cause of cancer related death in Mongolia. Early diagnosis is the very important management to increase successful treatment and survival rate. Glypican-3 (GPC3) protein is highly expressed in hepatocellular carcinoma (HCC) tissue and in serum of HCC patients. Recent studies have been conducted and suggested as a diagnostic biomarker for detecting HCC in the early stage. Therefore, we investigated the diagnostic value of the serum GPC3 level and compared it to the alpha-fetoprotein (AFP) level as a diagnostic biomarker of HCC.@*Methods@#We enrolled a total of 90 participants and divided into 3 groups with HCC (30), with liver cirrhosis (LC/30) and healthy (30) as the control group (30). GPC3 and AFP serum (sGPC-3, sAFP) levels were measured using commercially available enzyme-linked immunosorbent assay kits. The diagnostic accuracy was analyzed using the receiver operating characteristics (ROC) curve and estimated sensitivity and specificity of each biomarker. @*Results@#sGPC3 was significantly elevated in the HCC group as compared to liver cirrhosis and healthy subjects (658±138.2 pg/ml, 378±25.5 pg/ml, 356.3±29 pg/ml) respectively. sGPC-3 sensitivity was 96.6% and specificity was 100%. The area under the ROC curve (AUC) for GPC3 was 0.999 (0.996- 1.0).</br> In comparison, the mean of AFP was significantly higher in HCC (16.9±11.7 ng/ml) than in LC (6.7±7.6 ng/ml) and in healthy subject (3.3±2.1 ng/ml) and AFP sensitivity was 43,3 %, specificity was 95 % with an AUC of 0.808 (0.696- 0.921). </br> The combination of GPC-3 with AFP achieved the highest sensitivity (97.1%) and specificity (97%).@*Conclusion@#Serum GPC3 has a higher sensitivity than AFP for the early diagnosis of HCC. Combination of two markers showed greatest diagnostic accuracy.
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Adoptive immunotherapy based on chimeric antigen receptor-modified T cells (CAR-T) is one of the most promising strategies to treat malignant tumors, but its application in solid tumors is still limited. Glypican-3 (GPC3) is a meaningful diagnostic, therapeutic, and prognostic biomarker for hepatocellular carcinoma (HCC). The second/third generation GPC3-targeted CAR-T cells are generated to treat HCC. In order to improve the therapeutic effect, we constructed a fourth-generation lentiviral vector to express GPC3 CAR, human interleukin-7 (IL-7) and CCL19. Then the lentiviral vector and packaging plasmids were co-transfected into HEK293T cells to generate CAR lentiviral particles. Human T lymphocyte cells were transduced with CAR lentiviral to develop the fourth-generation GPC3-targeted CAR-T cells (GPC3-BBZ-7×19). In vitro, we used cell counting, transwell assay, luciferase bioluminescence assay and flow cytometry to compare the proliferation, chemotaxis, cytotoxicity and subtype distribution between GPC3-BBZ-7×19 CAR-T cells and the second generation GPC3-targeted CAR-T cells (GPC3-BBZ). In vivo, we established GPC3-positive HCC xenograft model in immunodeficient mice, then untransduced T cells (non-CAR-T) or GPC3-BBZ-7×19 CAR-T cells were injected. Tumor growth in mice was observed by bioluminescence imaging. Results showed that compared with GPC3-BBZ CAR-T, GPC3-BBZ-7×19 CAR-T cells had stronger proliferation, chemotactic ability, and higher composition of memory stem T cells (Tscm) (P values<0.05). However, there were no significant difference in cytotoxicity and cytokine secretion between them. In addition, GPC3-BBZ-7×19 CAR-T cells could significantly eliminate GPC3-positive HCC xenografts established in immunodeficient mice. Therefore, the fourth-generation GPC3-targeted CAR-T cells (secreting IL-7 and CCL19) are expected to be more durable and effective against HCC and produce tumor-specific memory, to provide a preclinical research basis for future clinical trials.
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Animals , Humans , Mice , Carcinoma, Hepatocellular , Cell Line, Tumor , Chemokine CCL19 , Metabolism , Glypicans , Metabolism , HEK293 Cells , Interleukin-7 , Metabolism , Lentivirus , Genetics , Liver Neoplasms , Receptors, Chimeric Antigen , Metabolism , T-Lymphocytes , Metabolism , Xenograft Model Antitumor AssaysABSTRACT
Aims: The main objective of this paper is to review the technologies used for the detection of Hepatocellular Carcinoma. Study Design: Convective cooling protects the cancer cells from thermal destruction and decreases the necrosed volume. A major objective of the method development is to achieve a virtually complete necrosis of tumors close to major blood vessels and to avoid blood vessel damage and, hence, the needed treatment planning. Place and Duration of Study: We found from this three-dimensional three-field coupling study that in large blood vessels, both convective cooling and acoustic streaming may change the temperature considerably near the blood vessel. Acoustic streaming velocity magnitude can be several times larger than the blood vessel velocity. Methodology: Different methods and techniques were proposed so far in the automatic detection of Hepatocellular Carcinoma. However the performance of the technologies till now not completely matched to the performance of human expert. Results: This review paper have analyzed the recent technologies in Liver Cancer Identification (24%), Liver Tumor Risks (16%), MR Imaging (22%), Liver Tumor Prevention (16%) and Liver Tumor Therapy (18%). The results presented in the current work can be further used to construct a surgical planning platform. Conclusions: Also we give some directions about the technologies and this can be useful for the researches to develop a new technology for the detection of Hepatocellular Carcinoma.
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Objective To investigate biological activity and antitumor effects of phosphatidylinositol-3 (GPC3) gene transfected dendritic cells(DC,DC-GPC3)co-cultured with cytokine induced killer cells(CIK)on hepatocellular carcinoma (HCC). Methods The phenotypes of effector cells were analyzed by flow cytometry. Levels of interleukin (IL)-2 and interferon (IFN)–γ were determined by MTT colorimetry and enzyme-linked immunosorbent assay, respectively. The cytotoxic activity against hepatocarcinoma cells (HepG2) was measured by lactate dehydrogenase release. Results The proportions of CD3+CD8+and CD3+CD56+double-positive cells were significantly elevated in the DCIK-GPC3 compared with the DCIK and CIK(P<0.05).Compared with the DCIK and CIK,the DCIK-GPC3 showed significantly higher levels of secreted IL-2 and IFN-γ in the supernatants(P<0.05).The antitumor effect of DCIK-GPC3 against HepG2 was the highest than that of DCIK and CIK at an effector-target ratio ranging from 20:1 to 50:1(P<0.05).Conclusion DCIK-GPC3 can enhance the cytotoxic activity against hepatocarcinoma cells in vitro.This study provides a theoretical and experimental basis for clinical immunotherapy using DCIK-GPC3.
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Objective To study the expression and significance of Glypican-3 in Budd-Chiari syndrome (BCS) complicated with hepatocellular carcinoma (HCC).Methods The data of 46 patients with BCS complicated with HCC (the BCS + HCC group) treated in The First Affiliated Hospital of Zhengzhou University from January 2007 to December 2016 were analyzed retrospectively.Another 48 patients with HBV-related HCC (the HBV + HCC group) and 43 patients with hepatic cyst (the hepatic cyst group) were randomly selected as the control groups during the same time period.The differencesin positive rates of Glypican-3 in the liver tissues among the three groups were compared.The BCS + HCC group was further divided into the Glypican-3 positive and Glypican-3 negative subgroups according to the expression of Glypican-3.The differences in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number,extrahepatic metastasis,vascular invasion,Edmondson-Steiner grading and BCLC staging between the two subgroups were compared.The survival time of the two subgroups was compared using the Kaplan-Meier method.Results The expression rates of Glypican-3 in the BCS + HCC group,HBV + HCC group and Hepatic Cyst group were 76.1％,70.8％ and 0％,respectively.The levels of Glypican-3 in the BCS + HCC group and the HCC group were significantly higher than that in the hepatic cyst group.The differences were statistically significant (P ＜ 0.05).No statistically significant difference was detected between the BCS + HCC group and the HBV + HCC group (P ＞ 0.05).In the group of patients with BCS + HCC,there was no significant difference in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number and extrahepatic metastasis between the Glypican-3 positive and negative subgroups (P ＞0.05).However,vascular invasion,Edmondson-Steiner grading and BCLC staging in the Glypican-3 positive subgroup were significantly higher than those in the Glypican-3 negative group,(P ＜ 0.05).The 1-year,3-year and 5-year survival rates were 77.1％,51.0％ and 22.8％ in the Glypican-3 positive subgroup,compared with 90.9％,63.6％ and 45.5％ in the Glypican-3 negative subgroup,respectively.There were statistically significant differences between the two groups (P ＜ 0.05).Conclusion Glypican-3 has a stable expression in patients with BCS complicated with HCC,and it is closely related to malignancy of the tumor and prognosis of the patients.
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Exosomes are small membrane bound vesicles typically 30 ～ 150 nm in size which are widely distributed in the body fluids.The cytosol is rich in proteins and nucleic acids which mediates communication between different cells and transporting substances.Exosomes are involved in pancreatic cancer initiation and progression by promoting the formation of tumor microenvironment,enhancing gemcitabine resistance.GPC1 + circulating exosomes and high exosome levels of microRNA-10b serve as potertial non-invasive diagnostic tools to detect pancreatic cancer in the early stage.Meanwhile,exosomes could successfully deliver genetic drugs,anticancer drugs to target cells.In this article,we provide a review on the recent advances on exosomes in the diagnosis and treatment of pancreatic cancer.
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Liver is the only organ in the human body that can regenerate to its original size after partial resection.Most studies focus on the mechanisms of the initiation and development of liver regeneration.Recently studies about termination of liver regeneration have been gradually investigating.Exploring the termination of liver regeneration can help us understand the mechanism of "hepatostat",and it can also help us discover the relationship between the destruction of liver regeneration termination process and tumorigenesis.This review will summarize the currently known signaling pathways in termination of liver regeneration.
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OBJECTIVE@#To investigate potential human leucocyte antigen (HLA)-A2-restricted epitope peptides of glypican-3 (GPC3) and determine the cytotoxicity of peptide-specific cytotoxic T lymphocytes (CTLs) against hepatocellular carcinoma (HCC) cells.@*METHODS@#The potential HLA-A*0201-restricted GPC3 peptides were screened using computer algorithms, T2 cell-binding affinity and stability of peptide/HLA-A*0201 complex assay. The peptide-specific CTLs were generated and their cytotoxicity against GPC3 SMMC 7721 and HepG2 cells was detected using IFN-γ based enzyme-linked immunospot and lactate dehydrogenase release assays in vitro.@*RESULTS@#A total of six peptides were identified for bindings to HAL-A2 and the GPC3 522-530 and GPC3 229-237 peptides with HLA-A*0201 molecules displayed high binding affinity and stability. The CTLs induced by the GPC3 522-530 or positive control GPC3 144-152 peptide responded to the peptide by producing IFN-γ, which were abrogated by treatment with anti-HLA-A2 antibody. The GPC3 522-530-specific CTLs responded to and killed SMMC 7721 and HepG2 cells, instead of GPC3-silenced SMMC 7721 or HepG2 cells. GPC3 522-530-specific CTLs response to HCC cells was blocked by anti-HLA-A2 antibody.@*CONCLUSIONS@#The GPC3 522-530 peptide contains antigen-determinant and its specific CTLs can effectively kill HCC in a HLA-A2-restricted and peptide-dependent manner. Our findings suggest that this peptide may be valuable for development of therapeutic vaccine.
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The aim of the experiments is to screen human single-chain variable fragment (scFv) targeting glypican 3 from phage display library, and analyze its biological activity. After several rounds of panning, the binders with high affinity were obtained through phage ELISA and IMGT analysis. The desired scFv gene was then ligated with pET-22b vector yielding recombinant plasmids, which was then introduced into E. coli Rosetta (DE3). Soluble scFv protein was expressed and further purified using Ni2+ affinity chromatography. The purified proteins were identified by SDS-PAGE and Western blot. Subsequently, the affinity and cell based binding activity were measured using Surface Plasmon Resonance (SPR) and flow cytometry assay, separately. Four enriched sequences with relatively high binding affinity were found (1F7, 1D7, 1D4 and 1B10). We also found that 1F7 scFv showed better targeting ability and higher affinity. The scFv could pave the way for new immunotherapies, such as bispecific anitbody, antibody-drug conjugate and chimeric antigen receptor T-cell immunotherapy cell, etc.
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Liver is the only organ in the human body that can regenerate to its original size after partial resection.Most studies focus on the mechanisms of the initiation and development of liver regeneration.Recently studies about termination of liver regeneration have been gradually investigating.Exploring the termination of liver regeneration can help us understand the mechanism of "hepatostat",and it can also help us discover the relationship between the destruction of liver regeneration termination process and tumorigenesis.This review will summarize the currently known signaling pathways in termination of liver regeneration.
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Objective To investigate potential human leucocyte antigen (HLA)-A2-restricted epitope peptides of glypican-3 (GPC3) and determine the cytotoxicity of peptide-specific cytotoxic T lymphocytes (CTLs) against hepatocellular carcinoma (HCC) cells. Methods The potential HLA-A*0201-restricted GPC3 peptides were screened using computer algorithms, T2 cell-binding affinity and stability of peptide/HLA-A*0201 complex assay. The peptide-specific CTLs were generated and their cytotoxicity against GPC3
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To assess the diagnostic utility of glypican-3 (GPC-3) in comparison to Hep Par 1 in the diagnosis of liver tumours, a cross-sectional study involving 66 resected liver tumours were tested for the protein expression of these markers by immunohistochemistry using monoclonal antibodies. Of the 66 cases, 26 (39.4%) were hepatocellular carcinoma (HCC), 4 (6.1%) were intrahepatic cholangiocarcinoma and 36 (54.5%) were metastatic tumours. Hep Par 1 and GPC-3 expressions in HCC were 24/26 (92.3%) and 19/26 (73.1%) respectively. In contrast, of non-HCC cases, only 2/40 cases (5.0%) expressed Hep Par 1, including a metastatic colorectal adenocarcinoma and a metastatic gastric adenocarcinoma. GPC-3 was expressed in 3/40 cases (7.5%), i.e. a metastatic adenocarcinoma of unknown origin, a metastatic gastric adenocarcinoma and an intrahepatic cholangiocarcinoma. The sensitivity and specificity for Hep Par 1 were 92.3% and 95% respectively while that of GPC-3 was 73.1% and 92.5% respectively. GPC-3 is a useful marker in the diagnosis of HCC. However it is not superior to Hep Par 1 in its sensitivity and specificity. We recommend that it is utilized together with Hep Par 1 as a panel in the diagnosis of HCC.
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BACKGROUND/AIMS: Glypican-3 (GPC3) protein is highly expressed in hepatocellular carcinoma (HCC) tissue. It has been suggested as a diagnostic biomarker, but its inconsistent performance means that it requires further assessment. We therefore investigated the diagnostic value of the plasma GPC3 level compared to the alpha-fetoprotein (AFP) level as a diagnostic biomarker of HCC. METHODS: We enrolled 157 consecutive patients with newly diagnosed HCC and 156 patients with liver cirrhosis (LC) as the control group. GPC3 plasma levels were measured using two commercially available enzyme-linked immunosorbent assays (ELISAs, named as Assay 1 and 2), and AFP levels were measured using an enzyme-linked chemiluminescent immunoassay. The diagnostic accuracy was analyzed using the receiver operating characteristics (ROC) curve. RESULTS: Plasma GPC3 levels in HCC patients were very low (0–3.09 ng/mL) in Assay 1, while only 3 of the 157 patients (1.9%) showed detectable GPC3 levels in Assay 2. The median GPC3 level was not significantly elevated in the HCC group (0.80 ng/mL) compared with the LC group (0.60 ng/mL). The area under the ROC curve (AUC) for GPC3 was 0.559 in Assay 1. In contrast, the median AFP level was significantly higher in HCC (27.72 ng/mL) than in LC (4.74 ng/mL), with an AUC of 0.729. CONCLUSION: The plasma level of GPC3 is a poor diagnostic marker for HCC, being far inferior to AFP. The development of a consistent detection system for the blood level of GPC3 is warranted.