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ABSTRACT BACKGROUND: Hepatocellular carcinoma (HCC) is the most frequent primary cancer of the liver and cirrhosis is considered a pre-malignant disease. In this context, the evolutionary sequence from low grade dysplastic nodule and high grade dysplastic nodule (HGDN) to early HCC and advanced HCC has been studied. The differential diagnosis between HGDN and early HCC is still a challenge, especially in needle biopsies OBJECTIVE: To evaluate an immunohistochemistry panel to differentiate dysplastic nodules and HCC. METHODS: Patients with cirrhosis who underwent surgical resection or liver transplantation were included. The sensitivity, specificity and accuracy for the diagnosis of neoplasia were analyzed by evaluating five markers: heat shock protein 70, glypican 3, glutamine synthetase, clathrin heavy chain and beta-catenin. P≤0.05 was considered statistically significant. RESULTS: One hundred and fifty-six nodules were included; of these, 57 were HCC, 14 HGDN, 18 low grade dysplastic nodules and 67 regenerative macronodules. Sensitivity of HCC diagnosis was 64.9% for glypican 3 and 77.2% for glutamine syntetase, while specificity was 96.0% and 96.0% respectively. When the panel of four markers was considered (excluding beta catenin), the specificity ranged from 87.9% for one positive marker to 100% for at least three markers. The best accuracy for HCC diagnosis was obtained with at least two positive markers, which was associated with a sensitivity of 82.5% and specificity of 99%. CONCLUSION: Differential diagnosis of dysplastic nodules and HCC by morphological criteria can be challenging. Immunomarkers are useful and should be used for the differential diagnosis between HCC and HGDN.
RESUMO CONTEXTO: O carcinoma hepatocelular (CHC) é o câncer primário do fígado mais frequente e a cirrose é considerada uma doença pré-maligna. Nesse contexto, a sequência evolutiva do nódulo displásico de baixo grau e nódulo displásico de alto grau (NDAG) para CHC precoce e CHC avançado tem sido estudada. O diagnóstico diferencial entre NDAG e CHC precoce ainda é um desafio, principalmente em biópsias por agulha. OBJETIVO: Avaliar um painel de imunohistoquímica para diferenciar nódulos displásicos de CHC. MÉTODOS: Foram incluídos pacientes com cirrose submetidos à ressecção cirúrgica ou transplante de fígado. A sensibilidade, especificidade e acurácia para o diagnóstico da neoplasia foram analisadas avaliando cinco marcadores: proteína de choque térmico 70kDa, glipican 3, glutamina sintetase, clatrina de cadeia pesada e beta-catenina. P≤0,05 foi considerado estatisticamente significativo. RESULTADOS: Cento e cinquenta e seis nódulos foram incluídos; destes, 57 eram CHC, 14 NDAG, 18 nódulos displásicos de baixo grau e 67 macronódulos regenerativos. A sensibilidade do diagnóstico de CHC foi de 64,9% para glipican 3 e 77,2% para glutamina sintetase, enquanto a especificidade foi de 96,0% e 96,0%, respectivamente. Quando o painel de quatro marcadores foi considerado (excluindo beta catenina), a especificidade variou de 87,9% para um marcador positivo a 100% para pelo menos três marcadores. A melhor acurácia para o diagnóstico de CHC foi obtida com pelo menos dois marcadores positivos, o que foi associado a uma sensibilidade de 82,5% e especificidade de 99%. CONCLUSÃO: O diagnóstico diferencial de nódulos displásicos e CHC por critérios morfológicos pode ser desafiador. Imunomarcadores são úteis e devem ser usados para o diagnóstico diferencial entre CHC e NDAG.
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Humans , Carcinoma, Hepatocellular/diagnosis , Neoplasms/diagnosis , Immunohistochemistry , Diagnosis, Differential , Liver Cirrhosis/diagnosisABSTRACT
Objective:To investigate the clinical and genetic characteristics of Simpson-Golabi-Behmel syndrome (SGBS) type Ⅰ caused by glypican-3 ( GPC3) gene mutations. Methods:Data of one neonate with SGBS type Ⅰ from Shenzhen Maternity and Child Healthcare Hospital Affiliated to Southern Medical University was reviewed retrospectively. Literature was retrieved to summarize the clinical and genetic characteristics of SGBS type Ⅰ caused by GPC3 mutations, using terms of "Simpson-Golabi-Behmel type Ⅰ", "GPC3" and "glypican-3" from China National Knowledge Infrastructure, VIP database, Wanfang database, and PubMed from January 2010 till April 2021. Results:The male infant was admitted to the hospital at 4 h after birth due to "abdominal distension for 1 h", presenting with dysmorphic facial features, including macrocephaly, coarse face, broad nasal bridge, macrostomia, tongue with a groove in the middle, as well as macrosomatia, supernumerary nipples, and hypospadias. Whole exome sequencing revealed a novel frameshift mutation (c.720delC) in GPC3 gene of the patient and his mother for hemizygous and heterozygous variation, respectively, based on which SGBS type Ⅰwas confirmed. During the follow-up, overgrowth, neuroblastoma, and motor development retardation were found in the boy. In addition to the index patient, 92 cases of SGBS type Ⅰ reported in 31 articles were analyzed, including 89(95.7%) males and 4(4.3%) females. The main clinical features were craniofacial dysmorphism, pre/postnatal overgrowth with multiple congenital anomalies. Most patients were combined with language disorders, motor retardation, and various degrees of dysnoesia, and were more likely to develop embryonic tumors. Among the 93 cases, 11(11.8%) suffered from tumors. Apart from 21 cases of termination, 63 cases were born alive and nine cases died after birth. Pathogenic variants in GPC3 gene were reported in 80 cases, which were nonsense mutation in 25 cases (31.2%), DNA fragment deletion in 21 cases (26.2%), frameshift mutation in 16 cases (20.0%), large duplications in eight cases (10.0%), missense mutation in five cases(6.2%), and splice site mutation in five cases(6.2%). Conclusions:SGBS type Ⅰ is an X-linked recessive genetic disease with various phenotypes. Patients with postnatal craniofacial dysmorphism, overgrowth, and multiple congenital anomalies should be highly suspected of SGBS type Ⅰ. Genetic testing is conducive to its early diagnosis. Treatment requires multidisciplinary cooperation and long-term follow-up, especially for those with tumors.
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Objective:To investigate the expression of glypican1 (GPC1) in pancreatic ductal adenocarcinoma (PDAC) and its relationship with the prognosis of patients with PDAC.Methods:From January 2015 to December 2018, 125 PDAC tumor specimens and corresponding para-carcinoma normal pancreatic tissue were collected from the Department of Pathology of Peking University Third Hospital. The expression of GPC1 protein was detected by the immunohistochemical Envision two-step method in all specimens. The specimens were divided into high and low GPC1 expression groups according to immunohistochemical scores, and the correlation between GPC1 protein expression and clinicopathological features and overall survival time was analyzed.Results:The positive expression rate of GPC1 protein was 0 score in 30.4% of PDAC tissues, 1 score in 15.2%, 2 score in 18.4% and 3 score in 36.0%, respectively, and high expression rate (2+ 3) was 54.4%. GPC1 protein was negatively expressed in para-carcinoma pancreatic tissues. The positive expression rate of GPC1 protein in PDAC tissue was significantly higher than that in para-carcinoma pancreatic tissue, and the difference was statistically significant ( P=0.000). The high expression of GPC1 protein was significantly correlated with tumor location and T stage ( P<0.05), but not with gender, age, history of diabetes and pancreatitis, preoperative blood CA19-9 level, postoperative surgical margin, tumor differentiation, lymph node metastasis, nerve invasion and vascular invasion (all P values >0.05). Univariate Cox proportional hazards analysis showed that GPC1 expression was associated with postoperative overall survival time in PDAC patients ( P<0.001). Multivariate Cox proportional hazards analysis showed that GPC1 protein expression level was an independent prognostic factor affecting overall survival time in PDAC patients ( P<0.001). The median survival time of PDAC patients with high GPC1 expression was significantly lower than that of PDAC patients with low GPC1 expression (11.00 months vs 18.00 months), and the difference was statistically significant ( P<0.01). Conclusions:GPC1 protein was abnormally high expressed in PDAC tumor tissue, and the high expression of GPC1 protein was positively correlated with tumor stage and negatively correlated with the overall survival time of patients. High expression of GPC1 was an independent risk factor for poor prognosis in PDAC patients.
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Phosphatidylinositol protein 3 (Glypican-3, GPC3) is a cell-surface heparan sulfate proteoglycan that belongs to the glycine related global membrane proteoglycan relatives.Many studies have shown that it is of great significance in the occurrence, diagnosis, treatment and prognosis of hepatocellular carcinoma (HCC).This paper reviews the recent studies on GPC3.
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Objective To investigate the upregulation of long-chain non-coding RNA LINC01204 on the expression of glypican-5 (GPC5) gene in bladder cancer cells and its effect on the proliferation,migration and invasion of bladder cancer cells.Methods Quantitative real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC01204 in 12 cases of bladder cancer tissues and paracancerous tissues,bladder cancer cell lines (BIU-87,T24,J82,5637) and normal bladder epithelial cell SV-HUC-1.The bladder cancer cells with the lowest level of LINC01204 were infected with lentivirus particles carrying LINC01204 (experimental group) or infected with negative control lentivirus particles (control group),and the expression of LINC01204 and GPC5 were detected by qRT-PCR in both groups of cells.Western blot was used to detect the expression of GPC5 protein in the two groups of cells.Cell counting kit-8 (CCK-8) and Transwell chamber assays were used to determine cell proliferation,migration and invasion.Results The expression of LINC01204 in bladder cancer tissues (0.773 ±0.063) was significantly higher than that in adjacent tissues (3.665 ± 0.330),with statistically significant difference (t =8.612,P ＜ 0.001).The expression of LINC01204 in bladder cancer cell lines BIU-87 (0.320 ± 0.034),T24 (0.515 ±0.056),J82 (0.644 ±0.039),and 5637 (0.147 ±0.018) were lower than that of normal bladder epithelial cells SV-HUC-1 (1.009 ± 0.077),with statistically significant difference (t =8.160,P＜0.001;t=5.179,P=0.002;t=4.221,P=0.006;t=10.890,P＜0.001).The expression of LINC01204 in 5637 cells was the lowest.The expression of LINC01204 and GPC5 mRNA in experimental group were significantly higher than that in the control group,with statistically significant difference [(11.000±1.028) vs (1.019 ±0.119),t =9.651,P＜0.001;(4.476 ±0.347) vs (1.046 ± 0.163),t =8.962,P ＜ 0.001],with statistically significant difference.Western blot showed that the expression of GPC5 protein was up-regulated.Compared with the control group,the proliferation ability of 5637 cells infected with LINC01204 in experimental group began to decrease significantly from the third day (0.686 ±0.044 vs 0.536 ±0.026,t =2.943,P =0.026).The number of migration cells in experimental group (118.300 ± 16.260) was significantly lower than that in the control group (208.200 ±22.930),with statistically significant difference (t =3.198,P =0.019).The number of cell invasion in experimental group (50.390 ±5.368) was significantly lower than that in the control group (97.480 ± 15.350),with statistically significant difference (t =2.896,P =0.028).Conclusions LINC01204 can upregulate the expression of GPC5 gene and inhibit the proliferation,migration and invasion of bladder cancer cells.Targeted therapy for LINC01204 is expected to become a new gene therapy method for bladder cancer.
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Objective: To construct the murine chimeric antigen receptor-redirected T lymphocyte (CAR-T) that can target glypican 3 (GPC3) and express interleukin-12 (IL-12) in an activation-dependent manner, and to explore the anti-cancer efficacy of CAR-T cells in immunocompetent mice bearing GPC3 positive (GPC3+) breast cancer. Methods: Gene recombination technology was used to construct the retroviral vector encoding GPC3-specific CAR (m9.28z) or encoding both GPC3-specific CAR and nuclear factor of activated T cells (NFAT)-IL-12 expression system (m9.28z/IL-12). Murine T cells were retrovirally transduced to develop m9.28z CAR-T cells and m9.28z/IL-12 CAR-T cells. In vitro, the cytotoxicity assay and ELISA assay were used to assess the biological functions of m9.28z/IL-12 CAR-T cells. In vivo, the GPC3+ breast cancer E0771-GPC3 cell subcutaneously transplanted model was established in C57BL/6 mice, and the tumor-bearing mice were injected with 5×106 murine untransduced T cells (mUTD, which served as the negative control), 5×106 m9.28z CAR-T cells, 2×106 m9.28z CAR-T cells, 5×106 m9.28z/IL-12 CAR-T cells and 2×106 m9.28z/IL-12 CAR-T cells, respectively. The growth of xenografts in nude mice was observed, and the expression levels of CD8α and forkhead box protein P3 (FOXP3) in tumor tissues were detected by immunohistochemistry. Results: The retroviral vector encoding m9.28z or m9.28z/IL-12 was constructed, and the m9.28z CAR-T cells and m9.28z/IL-12 CAR-T cells were developed subsequently. In vitro, the m9.28z/IL-12 CAR-T cells killed GPC3+ breast cancer cells specifically and effectively, and secreted more tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) than m9.28z CAR-T cells (both P < 0.05). In vivo, as compared with m9.28z CAR-T cells, the m9.28z/ IL-12 CAR-T cells suppressed the growth of xenografts significantly (P < 0.001), and there were more CD8+ T cells and less regulatory T cells (Treg) in tumor tissues treated with m9.28z/IL-12 CAR-T cells (both P < 0.01). Conclusion: The m9.28z/IL-12 CAR-T cells, which can express IL-12 inducibly, can display more potent anti-cancer activity against GPC3+ tumor than m9.28z CAR-T cells in vitro and in immunocompetent breast cancer transplanted mice.
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BACKGROUND/AIMS: Recent studies have suggested an important role of adipokines in the development of insulin resistance and diabetes mellitus. The clinical relevance of adipokines on long-term outcomes in patients with diabetes and chronic kidney disease is uncertain. The purpose of this study was to identify a predictable factor in patients with long-term diabetic complications. METHODS: A total of 161 diabetic individuals were followed-up from 2002 to 2013. Circulating plasma levels of adiponectin, glypican-4, irisin, visfatin, and visit-to-visit glucose variability were measured in diabetic patients. Associations among adipokines and variable metabolic parameters and microvascular, and macrovascular complications were evaluated. RESULTS: Plasma adiponectin and glypican-4 levels were significantly increased in patients with renal insufficiency. These adipokines were negatively associated with estimated glomerular filtration rate and positively associated with urinary albumin excretion. The relative risk of renal progression to dialysis increased independently with increasing level of adiponectin. Glypican-4 and visfatin were not predictive of any microvascular or macrovascular complications. Glucose variability increased the risk of diabetic nephropathy and cerebrovascular complications. CONCLUSIONS: Adiponectin and glypican-4 were associated with renal function and might be able to predict renal progression. Glucose variability was a predictable factor for diabetic nephropathy and cerebrovascular complications.
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Humans , Adipokines , Adiponectin , Diabetes Complications , Diabetes Mellitus , Diabetic Nephropathies , Dialysis , Glomerular Filtration Rate , Glucose , Glypicans , Insulin Resistance , Nicotinamide Phosphoribosyltransferase , Plasma , Renal Insufficiency , Renal Insufficiency, ChronicABSTRACT
Objective To evaluate the cytotoxicity of natural killer(NK)-92 cell lines against various human hepatocellular carcinoma cells(HCCs)and to explore the potential mechanism.Methods We established a culture method of NK-92 cell lines in vitro.Lactate debydrogenase(LDH)cytotoxicity assays and cytokine release assays were performed to determine whether NK-92 cell lines could recognize and kill HCCs in vitro.At the same time,Nu/Nu mices were housed. Subcutaneous(sc)xenografts HepG2 models of human hepatocellular carcinoma were established.1×107NK-92 cells were intravenously(iv)injected through the tail vein on days 2,9,16,23 while the control group was injected with PBS in the same way.Tumor size, tumor volume, tumor mass and mouse survival status were closely observed in experimental and control groups.Mice were euthanized when tumor-bearing time reached 28 days.Xenograft tissues were taken for general observation.Sections were cut and processed for HE staining and immunofluorescence staining.The expression of glypican-3(GPC3)protein in xenografts tissue was clearly defined.Results NK-92 cell lines that were chronically cultured in vitro and maintained typical phenotypic characteristics of NK cells with good cellular activity.Enhanced cytotoxicity and IFN-γ production of NK-92 cell lines were identified by LDH and ELISA,indicating that NK-92 cell lines could recognize and kill different kinds of HCCs.In addition,NK-92 cell lines efficiently suppressed the growth of HCC xenografts in vivo.Tumor volume in experimental group was significantly reduced compared with control group and there was low a GPC 3 expression in experimental group through immunofluorescence and immunohistochemistry results, pointing to the possibility that the cytotoxicity of NK cells was correlated with GPC3 +HCCs.Conclusion NK cells provide a promising means of therapeutic intervention for HCCs.NK-92 cell lines could eliminate HCC cells in vitro and in vivo.The cytotoxicity of NK-92 cell lines may work by killing the GPC3-positive cells in the liver cancer tissue.In addition to the anti-tumor effect, NK cells also have cytotoxicity on pathogens such as bacteria and viruses.
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Objective To investigate whether immunotherapy with dendritic cells (DC) transduced with glypican 3 (GPC3) (DC-GPC3) and cocultured with cytokine-induced killer cells (CIK) (DCIK-GPC3) is capable of inhibiting hepatocellular carcinoma (HCC) in vivo. Methods The hepatocarcinoma cell-bearing mouse models were established and divided randomly into four groups (A, B, C, D) for the injection of normal saline (NS), CIK, DC-CIK, and DCIK-GPC 3, respectively. Changes of tumor size, tumor inhibitory rate and cell apoptosis were compared between four groups. Results After treatment, the tumor volumes and weights were significantly decreased in D group compared with those of A, B and C groups (P<0.01). The inhibition rate was significantly increased in D group compared with that of B group and C group (P<0.01). The apoptotic rate was significantly increased in D group compared with that of A group, B group and C group (P<0.01). Conclusion DCIK-GPC3 can significantly inhibit tumor growth in vivo.
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Liver cancer is one of the most common malignant tumors in the world, and traditional liver cancer treatment methods have their own limitations.Glypican-3 (GPC3) is a cell-surface heparan sulfate proteoglycan and is involved in the regulation of individual development and cell proliferation and differentiation.It is also a hepatoma-specific carcinoembryonic antigen.The mechanism of action of GPC3 in the development and progression of liver cancer has become a hot research topic.GPC3 not only has a unique value in the diagnosis of liver cancer, but also plays an important role in the treatment of liver cancer.This article also introduces the application of GPC3-derived tumor vaccines, GPC3 antibodies, GPC3 gene therapy, and targeted therapy and brings new ideas for the treatment of liver cancer.
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Objective To investigate whether immunotherapy with dendritic cells (DC) transduced with glypican 3 (GPC3) (DC-GPC3) and cocultured with cytokine-induced killer cells (CIK) (DCIK-GPC3) is capable of inhibiting hepatocellular carcinoma (HCC) in vivo. Methods The hepatocarcinoma cell-bearing mouse models were established and divided randomly into four groups (A, B, C, D) for the injection of normal saline (NS), CIK, DC-CIK, and DCIK-GPC 3, respectively. Changes of tumor size, tumor inhibitory rate and cell apoptosis were compared between four groups. Results After treatment, the tumor volumes and weights were significantly decreased in D group compared with those of A, B and C groups (P<0.01). The inhibition rate was significantly increased in D group compared with that of B group and C group (P<0.01). The apoptotic rate was significantly increased in D group compared with that of A group, B group and C group (P<0.01). Conclusion DCIK-GPC3 can significantly inhibit tumor growth in vivo.
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[Summary] To evaluate the relationships between plasma GPC4 levels and HbA1C . Plasma GPC4 levels were measured in subjects with different glucose tolerances. The relationship between plasma GPC4 levels and HbA1C were studied. Increasing levels of GPC4 showed a significant linear trend and were independently associated with HbA1C . GPC4 levels gradually declined after gradually increased , which is similar with Fins, 2hIns, and AUCinsulin . GPC4 may be involved in the regulation of blood glucose and GPC4 may be regulated by insulin.
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Objective To explore the diagnostic and differential diagnostic value of Ki-67,Arg-1,HepPar-1 and Glypican-3 in primary hepatocellular carcinoma and benign hepatocellular lesions.Methods A total of 118 cases of primary hepatocellular carcinomas,100 cases of benign hepatocellular lesions and 263 cases malignant carcinomas originated from other organs were adopted in this study.Immunohistochemistry of Ki-67,Arg-1,HepPar-1 and Gly-3 were performed and evaluated in all specimens.Results The expressions of Ki-67 were concordant with the differentiation of hepatocellular carcinomas and the difference between primary hepatocellular carcinomas and benign hepatocellular lesions were statistically significant (P ＜0.01).The combination of Arg-1 + HepPar-1 + Glypican-3 had the most sensitivity and specificity in differential diagnosis between primary hepatocellular and metastatic malignant cancers,while the combination of Ki-67 + Glypican-3 + Arg-1 showed the better diagnostic value in primary hepatocellular carcinomas and benign lesions.Conclusions The application of different immunohistochemical antibody spectra in primary liver cancers could efficiently improve diagnostic accuracy and reliability.
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BACKGROUND: Previous studies have reported that glypican-4 (GPC4) regulates insulin signaling by interacting with insulin receptor and through adipocyte differentiation. However, GPC4 has not been studied with regard to its effects on clinical factors in patients with type 2 diabetes mellitus (T2DM). We aimed to identify factors associated with GPC4 level in T2DM. METHODS: Between January 2010 and December 2013, we selected 152 subjects with T2DM and collected serum and plasma into tubes pretreated with aprotinin and dipeptidyl peptidase-4 inhibitor to preserve active gastric inhibitory polypeptide (GIP) and glucagon-like peptide 1 (GLP-1). GPC4, active GLP-1, active GIP, and other factors were measured in these plasma samples. We performed a linear regression analysis to identify factors associated with GPC4 level. RESULTS: The subjects had a mean age of 58.1 years, were mildly obese (mean body mass index [BMI], 26.1 kg/m2), had T2DM of long-duration (mean, 101.3 months), glycated hemoglobin 7.5%, low insulin secretion, and low insulin resistance (mean homeostatic model assessment of insulin resistance [HOMA-IR], 1.2). Their mean GPC4 was 2.0±0.2 ng/mL. In multivariate analysis, GPC4 was independently associated with age (β=0.224, P=0.009), and levels of active GLP-1 (β=0.171, P=0.049) and aspartate aminotransferase (AST; β=–0.176, P=0.043) after being adjusted for other clinical factors. CONCLUSION: GPC4 was independently associated with age, active GLP-1, and AST in T2DM patients, but was not associated with HOMA-IR and BMI, which are well known factors related to GPC4. Further study is needed to identify the mechanisms of the association between GPC4 and basal active GLP-1 levels.
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Humans , Adipocytes , Aprotinin , Aspartate Aminotransferases , Body Mass Index , Diabetes Mellitus , Diabetes Mellitus, Type 2 , Gastric Inhibitory Polypeptide , Glucagon-Like Peptide 1 , Glypicans , Glycated Hemoglobin , Insulin , Insulin Resistance , Linear Models , Multivariate Analysis , Plasma , Receptor, InsulinABSTRACT
Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen glypican-3 (GPC3) and its preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. co-liBL21. Spleen cells were obtained from Balb/c mice embedding immunized with purified antigen intraperitoneally, and fused with Sp2/0 cells. Hybridoma cells were screened by indirect ELISA, and identified by Western blot assay using puri-fied protein after the cell fusion. The indirect immunofluorescence method was used to detect the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stably expressed in E. coliBL21. A mouse hybridoma cell line secreting monoclonal antibody to GPC3 was obtained. Western blot analysis showed that monoclonal antibody specifically recognized recombinant protein. Monoclonal antibody could be used to detect GPC3 protein expression in HepG2 cell line by indirect immunofluorescence. Conclusion The monoclonal antibody against GPC3 was successfully obtained.
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Objective To evaluate the clinical significance of combined detection of phosphatidyhnositol 3 proteoglycans (GPC3) and alpha-fetoprotein heterogeneity (AFP-L3) and total bile acids (TBA) in diagnosis of primary hepatocellular carcinoma.Methods Collected 154 cases of primary hepatocellular carcinoma patients (hepatocellular carcinoma group) and 78 cases of cirrhosis patients (cirrhosis group) and 56 normal controls (control group) from May 2011 to December 2012.The level of GPC3,AFP-L3 was measured by enzyme-linked immunosorbent assay,the level of alpha-fetoprotein (AFP) was determined by mdioimmunoassay,and the level of TBA was measured by enzymatic cycle,and they were compared.Results The level of GPC3,AFP-L3 and TBA was (10.70 ± 3.10) μ g/L,(338.60 ± 379.20) μ g/L,(79.91 ± 70.64) μ mol/L in hepatocellular carcinoma group,which was higher than that in cirrhosis group [(2.70 ±0.71) μg/L,(6.45 ±2.79) μg/L,(33.10 ±21.90) μmol/L] and control group [(1.28 ± 0.60) μ g/L,(0.68 ± 0.56) μ g/L,(5.40 ± 2.20) μ mol/L],the level of GPC3,AFP-L3 and TBA in cirrhosis group was higher than that in control group,and there was significant difference (P＜ 0.01).The sensitivity of three tumor markers combined detection was 97.2％ (280/288),which was significantly higher than that of individual detection of GPC3 (72.1％,111/154),AFP-L3 (53.2％,82/154),TBA (94.8％,146/154),the difference was statistically significant (P ＜ 0.01).Conclusion Serum GPC3,AFP-L3 and TBA combined detection can improve the sensitivity of primary hepatocellular carcinoma and has important chnical significance in early diagnosis of primary hepatocellular carcinoma.
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Objective To investigate the expression characteristics of Glypican3 ,MMP-9 and MMP-14 in primary hepatocellular carcinoma ,and to focus on their roles on the development ,progress and metastasis of tumor .Methods 102 cases of primary hepato-cellular carcinoma were taken as the observation group and 80 cases of normal liver tissues as the control group .The expressions of Glypican3 ,MMP-9 and MMP-14 were detected by the immunohistochemistry method and their expression difference in different clinicopathological characteristics and the correlation were investigated .Results The positive rates of Glypican3 ,MMP-9 and MMP-14 expressions in the observation group were significantly higher than those in the control group .The positive rates of Glypi-can3 ,MMP-9 and MMP-14 expressions were closely correlated with the tumor volume ,differentiation degree ,lymph node metasta-sis ,vessel infiltration ,membrane invasion and PCNA expression .The correlation analysis showed that the positive relationships were found between Glypican3 and MMP-9 ,between Glypican3 and MMP-14 and between MMP-9 and MMP-14 in the observation group(r=0 .48 ,P=0 .024 1 ;r=0 .46 ,P=0 .013 2;r=0 .43 .P=0 .031 3) .The survival analysis showed that expressions of Glypi-can3 ,MMP-9 and MMP-14 were correlated with the patient′s prognosis .Conclusion The higher-expressions of Glypican3 ,MMP-9 and MMP-14 may promote the occurrence and development of primary hepatocellular carcinoma .Detecting the postoperative expres-sions of Glypican3 ,MMP-9 and MMP-14 has certqain value to judge the prognosis in primary hepatocellular carcinoma .
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Objective To explore the value of arginase-1(Arg-1) and glypican-3 (GPC-3) combined examination in the differential diagnosis of hepatocellular carcinoma (HCC),metastatic carcinoma (MC) of liver and intrahepatic cholangiocarcinoma (ICC).Methods From January 2005 to December 2011,a total of 54 patients with HCC were selected,including 10 cases with high differentiation,25 cases with moderate differentiation and 19 cases with poor differentiation.At the same time,25 patients with MC of liver and 20 patients with ICC were selected.A total of 31 normal liver specimens were set as control.The expressions of Arg 1 and GPC-3 in the above tissues were detected by immunohistochemistry method.The sensitivity and specificity of the examination in the diagnosis of HCC were analyzed.Chi-square test was performed for count data analysis.Results The positive expression rate of Arg-1 in HCC,MC of liver,ICC and normal liver tissues was 87.0％ (47/54),4.0％ (1/25),5.0％ (1/20) and 100.0％ (31/31),respectively.The Arg 1 positive expression rate in HCC tissues was higher than that in other tumor tissues of non-HCC,and the difference was statistically significant (x2 =66.98,P＜0.05).The positive expression rate of GPC-3 in HCC,MC of liver,ICC and normal liver tissues was 70.4％ (38/54),12.0％ (3/25),5.0％ (1/20) and 0 (0/31),respectively.The GPC-3 positive expression rate in HCC tissues was higher than that in other tumor tissues of non-HCC and the difference was statistically significant (x2=37.98,P＜0.05).The sensitivity and specificity of Arg-1 or GPC-3 positive in HCC diagnosis was 92.6％ (50/54) and 86.7％ (39/45).The sensitivity and specificity of both Arg 1 and GPC-3 positive in HCCdiagnosis was 64.8％ (35/54) and 100.0％ (45/45).Conclusion Arginase-1 and glypican-3 combined examination has an important value in HCC diagnosis and differential diagnosis.
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Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.
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Objective: To evaluate the feasibility of using anti-GPC3 monoclonal antibody in discriminating benign and malignant hepatocellular lesions. Methods: The expression of GPC3 was examined by semiquantitative immunohistochemistry assessment in 19 normal control samples, 94 hepatocellular carcinoma samples, 20 intrahepatic cholangiocarcinoma samples and 42 benign hepatocellular samples. The relation of GPC3 expression profile with the clinicopathological factors of HCC was analyzed. Results: GPC3 staining was positive in 75 (79.8%) of the 94 HCC samples. And GPC3 expression was not detected in normal liver tissue adjacent to hepatic hemangioma, intrahepatic cholangiocarcinoma, or benign hepatic lesions. GPC3 protein expression in HCCs was correlated with serum alpha-fetoprotein level (P = 0.020), tumor encapsulation (P = 0.043), and portal vein invasion (P = 0.050). Conclusion: Anti-human GPC3 mouse monoclonal antibody can specifically detect GPC3 protein expression in hepatocellular carcinomas tissues, which warrants further examination for future clinical application.