Research Progress on Glypican-3 Targeted Therapy in Hepatocellular Carcinoma / 药学实践杂志

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.

Fluorescence imaging in hepatocellular carcinoma patients by using a glypican 3-specific probe / 中华肝胆外科杂志

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.

A new diagnostic biomarker in early detection of Hepatocellular Carcinoma / Монголын Анагаах Ухаан

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.

Effect of AU-rich element RNA-binding factor 1 on the level of glypican 3 in hepatocellular carcinoma / 临床肝胆病杂志

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.

Construction and function of Glypican-3-targeted fourth-generation chimeric antigen receptor T cells (secreting IL-7 and CCL19) / 生物工程学报

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.

The expression and significance of Glypican-3 in Budd-Chiari syndrome complicated with hepatocellular carcinoma / 中华肝胆外科杂志

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.

Research progress in molecular mechanisms in termination of liver regeneration / 中华肝胆外科杂志

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.

Research progress in molecular mechanisms in termination of liver regeneration / 中华肝胆外科杂志

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.

Screening and characterization of human anti-GPC3 single chain Fv antibody fragment selected by phage display / 药学学报

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.

Glypican-3-specific cytotoxic T lymphocytes induced by human leucocyte antigen-A*0201-restricted peptide effectively kill hepatocellular carcinoma cells in vitro

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

Glypican-3-specific cytotoxic T lymphocytes induced by human leucocyte antigen-A*0201-restricted peptide effectively kill hepatocellular carcinoma cells in vitro

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.

Glypican-3 is useful but not superior to Hep Par 1 in differentiating hepatocellular carcinoma from other liver tumours

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.

Research progress of a new therapeutic target in hepatocellular carcinoma glypican-3 / 中国药理学通报

Glypican-3 ( GPC3 ) plays very important role in the regulation of cell growth and differentiation in hepatocellular car-cinoma ( HCC ) . GPC3 is closely related to the occurrence and development of HCC. A dramatic elevation of GPC3 expression has been reported in a large proportion of HCC, which suggests that GPC3 is remarkably sensitive and specific to the diagnosis of HCC. GPC3 is a potential therapeutic target of HCC. This paper reviews the structure and function of GPC3, the progress of im-munotherapy based on GPC3 of HCC, and discusses the prospect of therapeutic target of liver cancer in the future.

Expression and Clinical Significance of Glypican-3 in Hepatocellular Carcinoma Tissues / 现代检验医学杂志

Objective To study the expression and the clinical significance of glyican-3 (GPC3)in hepatocellular carcinoma (HCC)tissues.Methods Immunohistochemical method was performed to evaluate the expression of GPC3 in 54 cases of HCC tissues,46 cases of para-carcinoma tissues,22 cases of cirrhosis tissues,12 cases of normal liver tissues,the correlation between expression levels of GPC3 and clinicopathologic factors was analyzed.Results GPC3 protein was not expressed in normal livers tissues,the levels of GPC3 (7.39±3.64)and the positive rate (81.48%)in HCC tissues were significantly higher than para-carcinoma tissues (1.15±0.99,13.04%),cirrhosis tissues (0.32±0.56,4.54%),the difference was sta-tistically significant (P 0.05);the positive rate in HCC tissues was independent of sex,age, serum HBsAg,alpha-fetoprotein (AFP),tumor diameter,metastasis,clinical stage (P >0.05),only related to the cirrhosis (P <0.05).The high expression rate of GPC3 in HCC tissues was correlated with AFP,tumor diameter,cirrhosis,metasta-sis,clinical stage (P <0.05).Conclusion GPC3 has important clinical significance in the diagnosis of HCC,the expression level of GPC3 associated with the progression of HCC.

Glypican-3 level assessed by the enzyme-linked immunosorbent assay is inferior to alpha-fetoprotein level for hepatocellular carcinoma diagnosis

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.

Analysis of the expression of AFP-L3 and GPC-3 in primary hepatic carcinoma with different concentration of AFP / 国际检验医学杂志

Objective To analyse the expression of alpha-fetoprotein variant-L3(AFP-L3)and glypican-3(GPC-3)in primary he-patic carcinoma(PHC)with different concentration of alpha-fetoprotein (AFP),so as to provide references for the diagnosis of PHC.Methods 240 cases of outpatients,inpatients and individuals on physical examination were selected as subjects and serum levels of AFP-L3 and GPC-3 were detected.All of the subjects were divided into negative group(0≤AFP<20 ng/mL),low concen-tration group(20≤AFP<400 ng/mL)and high concentration group(AFP≥400 ng/mL)according to the serum levels of AFP.Ser-um levels of AFP-L3 and GPC-3 were compared among the three groups.And the sensitivity,specificity and accuracy of single de-tection of AFP-L3 or GPC-3 and those of combined detection of AFP-L3 and GPC-3 were compared as well.Results The serum levels of AFP-L3 and GPC-3 in the low concentration group and high concentration group were both higher than those in the nega-tive group,and those in the high concentration group were also higher than those in the low concentration group,had statistically significant differences(P < 0.05 ).The sensitivity,specificity and accuracy of combined detection of AFP-L3 and GPC-3 were 84.4%,95.9% and 93.8% respectively,which were higher than those of single detection of AFP-L3 or GPC-3.Conclusion Com-bined detection of AFP-L3 and GPC-3 could improve the sensitivity,specificity and accuracy for diagnosis of PHC,which has clinical significance for the diagnosis of PHC.

Diagnostic utility of immunohistochemical markers SALL4, D2-40 and Glypican-3 in primary testicular germ cell tumors / 临床与实验病理学杂志

Purpose To investigate the diagnostic utility of the immunohistochemical markers SALL4, D2-40 and Glypican-3 in prima-ry testicular germ cell tumors (TGCTs). Methods The expression of SALL4, D2-40 and Glypican-3 protein was detected by EnVi-sion immunohistochemical method in 56 cases of primary testicular germ cell tumors, including 5 intratubular germ cell neoplasms ( IT-GCNs) , 10 seminomas, 14 embryonal carcinomas ( ECs) , 14 yolk sac tumors ( YSTs) , 1 choriocarcinoma, 5 immature teratomas and 12 mature teratomas. 10 normal testicular tissues and 5 lymphomas were selected as control. Results All of ITGCNs, seminomas, YSTs and ECs were diffusely strongly positive for SALL4. Focal SALL4 staining was seen in choriocarcinoma, 3 of 5 immature terato-mas and 3 of 12 mature teratomas. All of ITGCNs, seminomas showed diffusely strong D2-40 staining. ECs (4/14) were focally posi-tive for D2-40, while choriocarcinoma, YSTs and teratomas were negative for D2-40. Glypican-3 was diffusely positive in YSTs (13/14), and focally weakly positive in ECs (2/14), respectively. ITGCNs, seminomas, choriocarcinoma and teratoma were negative for Glypican-3. In contrast, 10 normal testicular tissues and 5 lymphomas showed no SALL4, D2-40 and Glypican-3 staining. Conclu-sions SALL4 is a useful diagnostic marker with high sensitivity and specificity for TGCTs. Combination of SALL4, D2-40 and Glypi-can-3 is helpful to the diagnosis and differential diagnosis for TGCTs.

Clinical Utility of Plasma Glypican-3 and Osteopontin as Biomarkers of Hepatocellular Carcinoma

BACKGROUND/AIMS: alpha-Fetoprotein (AFP) is the biomarker most widely used to detect hepatocellular carcinoma (HCC), despite its suboptimal diagnostic accuracy. Glypican-3 (GPC3) and osteopontin (OPN) are secreted glycoproteins that are reportedly associated with tumorigenesis and metastasis. This study was conducted to evaluate the clinical utility of using plasma GPC3 and OPN as diagnostic biomarkers for HCC. METHODS: We measured the plasma levels of GPC3 and OPN in 120 HCC and 40 chronic liver disease (CLD) patients via an enzyme-linked immunosorbent assay. The diagnostic accuracy of each tumor marker was evaluated using receiver operating characteristic (ROC) curve analysis. RESULTS: The GPC3 levels in the HCC patients (75.8 ng/mL) were significantly higher (p=0.020) than the levels in patients with CLD (66.4 ng/mL). The area under the ROC curve (AUROC) values for GPC3 and OPN were 0.62 and 0.51, respectively. In subgroup analyses, including subgroups of HCC patients with low serum AFP and PIVKA II levels, the AUROC of GPC3 remained relatively high (0.66), and GPC3 showed a high sensitivity (62.1%) for detecting small HCC tumors. CONCLUSIONS: The plasma levels of GPC3 and OPN demonstrated low diagnostic accuracy for HCC. However, GPC3 may have a complementary role in diagnosing HCC in patients with nondiagnostic levels of conventional tumor markers and with small-sized tumors.

Cytotoxic effects of antiglypican-3 against HepG2 cell line.

For decades, various systemic therapies have been explored for the treatment of advanced hepatocellular carcinoma (HCC), the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. Nevertheless, no satisfactory results have been obtained so far. Glypican-3 (GPC-3) is a cell-surface heparan sulfate proteoglycans (HSPGs) that emerged as a promising diagnostic marker as well as target for therapy. Therefore; we investigated antitumor activity of antiglypican-3 (antiGPC3), a specific antibody aginst GPC-3, against HepG2, human HCC, cell line. HepG2 cells were treated with AntiGPC3 at (5, 10 and 20 μg/ml). HepG2 cell proliferation was measured by MTT and lactate dehydrogenase (LDH) assays. GPC-3, HSPG and sulfatase-2 (SULF2) levels were measured by ELISA. Moreover, apoptosis was assessed by measuring Caspase-3 activity. We found that, antiGPC3 reduced HepG2 cells survival and showed cell cytotoxicity in a dose-dependent manner. In addition, antiGPC3 was able to increase the apoptosis measured by capase-3 activity in hepG2 cells. Finally, antiGPC3 restored HSPG level without affecting SULF2 in HepG2 cells. We can conclude that, antiGPC3 possesses cytotoxic effects, which can be partially explained by restoration of HSPGs and increase of caspase-3 apoptotic pathway. GPC-3 represents a promising target of HCC therapy.

Glypican-3 as a tumor marker for hepatocellular carcinoma.

Hepatocellular carcinoma(HCC) is one of the most frequent cancers in the world.The burden of HCC has been increasing in Egypt with a doubling in the incidence in the past ten years. The prognosis of most patients is unsatisfactory due to rapid clinical deterioration after the initial diagnosis. Therefore, it is very important to detect HCC and the recurrence at its earlier period. Glypican-3(GPC3) is a cell-surface protein, which is a member of the heparan sulfate proteoglycan family. GPC3 was indentified in the serum of patients with HCC and can be used as a serological test for the diagnosis of HCC. The aim was to assess the value of serum GPC3 in Egyptian patients with HCC. This study was included 30 patients with HCC, 30 patients with liver cirrhosis and 20 healthy controls. For all groups we studied clinical data, image findings, serum alphafetoprotein(AFP) levels detected by enzyme immunoassay(EIA) kit and GPC3 gene expression was detected by real time polymerase chain reaction(PCR).Tumour characteristics were assessed including size, number and site. Tumour staging was done using Okuda &Tokyo staging systems.The data showed that HCC patients had a significantly higher mean GPC3 values (p=0.000)than both cirrhotics and healthy control groups. GPC3 has a positive significant correlation with tumour size (p=0.015)and Tokyo staging system(p=0.047).GPC3 could be a useful diagnostic & prognostic marker for detection of HCC.