IM To determine whether antisense insulinlike growth factorI(IGFI) gene can modulate CEA and AFP expression in human hepatoma cells (HepG2).METHODS Transfection of HepG2 cells was accomplished using Lipofectin reage...IM To determine whether antisense insulinlike growth factorI(IGFI) gene can modulate CEA and AFP expression in human hepatoma cells (HepG2).METHODS Transfection of HepG2 cells was accomplished using Lipofectin reagent. Northern blot analysis confirmed the antisense IGFI RNA of the transfected cells. CEA and AFP levels were measured using radioimmunoassay.RESULTS Human hepatoma cell lines (HepG2) were transfected with antisense IGFI gene. Northern blot analysis confirmed that antisense IGFI RNA was expressed in the transfected cells. The effect of antisense IGFI gene on CEA and AFP expression was demonstrated by the fact that the CEA and AFP levels in the supernatant of transfected cell culture were significantly lower as compared with the parent cells, 〔CEA 70μg/L±076μg/L and 329μg/L±180μg/L (P<005) and AFP 5363μg/L±602μg/L and 90μg/L±526μg/L (P<001), respectively〕.CONCLUSION The malignant potentiality of the transfected cells was partially suppressed. Antisense IGFI gene can modulate the expression of CEA and AFP in human hepatoma cell lines (HepG2).展开更多
Direct acting antiviral(DAA)treatments may reduce the elevatedαfetoprotein(AFP),but data on how these treatments affect elevated AFP in patients with chronic hepatitis C(CHC)remain insufficient.In the present study,t...Direct acting antiviral(DAA)treatments may reduce the elevatedαfetoprotein(AFP),but data on how these treatments affect elevated AFP in patients with chronic hepatitis C(CHC)remain insufficient.In the present study,the frequency of baseline AFP elevations and their related factors,AFP dynamics during and after DAA treatment,and factors associated with AFP reduction was assessed.This retrospective study included 141 patients with CHC without hepatocellular carcinoma who received DAA and achieved sustained virological response.The details are as follows:mean post-treatment follow-up was 99 weeks(12–213);mean age,57.8 years old;52%,males;79%,genotype(GT)1;and 47%,cirrhosis.Pre-treatment AFP elevation(>5.5 ng/mL)was seen in 48.2%patients.On multivariate analysis,baseline AFP>5.5 was associated with the presence of cirrhosis(P=0.001),co-existing non-alcoholic steatohepatitis(NASH)(P=0.035),and GT 1(P=0.029).AFP normalization was seen in 28.2%patients at treatment week 2,in 52%at the end of treatment,and in 73.4%at the end of follow-up.Post-treatment week 24 AFP normalization was associated with the absence of cirrhosis(P=0.003),Child--Pugh score<6(P=0.015),and baseline AFP<10(P=0.015).AFP elevation is common in patients with CHC and independently associated with NASH,cirrhosis,and GT 1.DAA treatment resulted in AFP normalization as early as treatment week 2.Post-treatment week 24 AFP normalization is independently associated with the absence of cirrhosis,Child--Pugh score<6,and baseline AFP<10.展开更多
To clone the murine α fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1 6 cells, and then the murine α fetoprotein gene was a...To clone the murine α fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1 6 cells, and then the murine α fetoprotein gene was amplified by RT PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. After transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8kb murine α fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.展开更多
Hepatocellular carcinoma(HCC) is the fifth most common cancer and the second leading cause of cancer-related deaths worldwide. Although the prognosis of patients with HCC is generally poor, the5-year survival rate is ...Hepatocellular carcinoma(HCC) is the fifth most common cancer and the second leading cause of cancer-related deaths worldwide. Although the prognosis of patients with HCC is generally poor, the5-year survival rate is > 70% if patients are diagnosed at an early stage. However, early diagnosis of HCC is complicated by the coexistence of inflammation and cirrhosis. Thus, novel biomarkers for the early diagnosis of HCC are required. Currently, the diagnosis of HCC without pathological correlation is achieved by analyzing serum α.fetoprotein levels combined with imaging techniques. Advances in genomics and proteomics platforms and biomarker assay techniques over the last decade have resulted in the identification of numerous novel biomarkers and have improved the diagnosis of HCC. The most promising biomarkers,such as glypican-3, osteopontin, Golgi protein-73 and nucleic acids including microRNAs, are most likely to become clinically validated in the near future. These biomarkers are not only useful for early diagnosis of HCC, but also provide insight into the mechanisms driving oncogenesis. In addition, such molecular insight creates the basis for the development of potentially more effective treatment strategies. In this article,we provide an overview of the biomarkers that are currently used for the early diagnosis of HCC.展开更多
AIM:To evaluate diagnostic value ofα-fetoprotein (AFP)-L3 and prothrombin induced by vitamin K absence-Ⅱ(PIVKA-Ⅱ)in hepatocellular carcinoma(HCC). METHODS:One hundred and sixty-eight patients during routine HCC sur...AIM:To evaluate diagnostic value ofα-fetoprotein (AFP)-L3 and prothrombin induced by vitamin K absence-Ⅱ(PIVKA-Ⅱ)in hepatocellular carcinoma(HCC). METHODS:One hundred and sixty-eight patients during routine HCC surveillance were included in this study.Of the 168 patients,90(53.6%)had HCC including newly developed HCC(n=82)or recurrent HCC after treatment(n=8).Sera were obtained during their first evaluation for HCC development and at the time of HCC diagnosis before commencing HCC treatment.HCC was diagnosed by histological examination,appropriate imaging characteristics-computed tomography or magnetic resonance imaging.Control sera were collected from 78 patients with benign liver disease(BLD),which were obtained during routine surveillance with a suspicion of HCC.AFP,AFP-L3 and PIVKA-Ⅱwere measured in the same serum by microchip capillary electrophoresis and liquid-phase binding assay on a micro-total analysis system Wako i30 auto analyzer.The performance characteristics of three tests and combined tests for the diagnosis of HCC were obtained using receiver operating characteristic curves in all populations and subgroups with AFP<20 ng/mL. RESULTS:Of 90 HCC patients,38(42.2%)patients had AFP<20 ng/mL,20(22.2%)patients had AFP 20-200 ng/mL and 32(35.6%)patients had AFP>200 ng/mL.Of the 78 BLD patients,74(94.9%)patients had AFP<20 ng/mL.After adjustment for age and HBV infection status,AFP-L3 levels were higher in HCC than in BLD among patients with low AFP levels(<20 ng/mL)(P<0.001).In a total of 168 patients,areas under the curve(AUC)for HCC were 0.879,0.887,0.801 and 0.939 for AFP,AFP-L3,PIVKA-Ⅱand the combined markers,respectively.The combined AUC for three markers showed higher value than the AUCs of individual marker(P<0.05).AFP-L3 had higher AUC value than PIVKA-Ⅱfor HCC detection in entire patients(P =0.043).With combination of AFP-L3(cut-off>5%) and PIVKA-Ⅱ(cut-off>40 AU/L),the sensitivity were 94.4%and specificity were 75.6%in all patients.In 112 patients with low AFP levels(<20 ng/mL),AUCs展开更多
AIM:To evaluate the diagnostic value of glypican-3(GPC3) in serum and liver for primary hepatocellular carcinoma(HCC).METHODS:Serum levels of GPC3 and α-fetoprotein(AFP) were measured in 75 patients with primary HCC ...AIM:To evaluate the diagnostic value of glypican-3(GPC3) in serum and liver for primary hepatocellular carcinoma(HCC).METHODS:Serum levels of GPC3 and α-fetoprotein(AFP) were measured in 75 patients with primary HCC and 32 patients with liver cirrhosis.Expression of GPC3 and AFP in 58 HCC and 12 cirrhotic specimens was detected with immunohistochemical staining.RESULTS:When the cut-off value of serum GPC3 was set at 300 ng/L,its sensitivity and specificity for HCC were 47.0% and 93.5%,respectively.Among the 14 patients with HCC at stage according to the Barcelona Clinic Liver Cancer staging system,the serum GPC3 level was higher than 300 ng/L in 50%(7/14) patients,the serum AFP level was not ≥ 400 μg/L in any patient.Combined serum AFP and GPC3 significantly increased the sensitivity to the diagnosis of HCC.The GPC3 expression was detected in cytoplasm of HCC cells but not in hepatocytes and bile ducts of benign tumors.Among the 58 HCC patients,the GPC3 was expressed in 100%(28/28) patients with their serum AFP level ≥ 400 μg/L,and in 90%(27/30) patients with their AFP level < 400 μg/L,respectively.The GPC3 was weakly or negatively expressed in all paracarcinomatous and cirrhotic tissue samples.AFP positive HCC cells were only found in 1 out of the 58 HCC patients.CONCLUSION:GPC3 protein is a sensitive and specific serum marker for diagnosis of early HCC.Its expression in liver tissues can be used to discriminate tumor cells from benign hepatic cells.展开更多
文摘IM To determine whether antisense insulinlike growth factorI(IGFI) gene can modulate CEA and AFP expression in human hepatoma cells (HepG2).METHODS Transfection of HepG2 cells was accomplished using Lipofectin reagent. Northern blot analysis confirmed the antisense IGFI RNA of the transfected cells. CEA and AFP levels were measured using radioimmunoassay.RESULTS Human hepatoma cell lines (HepG2) were transfected with antisense IGFI gene. Northern blot analysis confirmed that antisense IGFI RNA was expressed in the transfected cells. The effect of antisense IGFI gene on CEA and AFP expression was demonstrated by the fact that the CEA and AFP levels in the supernatant of transfected cell culture were significantly lower as compared with the parent cells, 〔CEA 70μg/L±076μg/L and 329μg/L±180μg/L (P<005) and AFP 5363μg/L±602μg/L and 90μg/L±526μg/L (P<001), respectively〕.CONCLUSION The malignant potentiality of the transfected cells was partially suppressed. Antisense IGFI gene can modulate the expression of CEA and AFP in human hepatoma cell lines (HepG2).
文摘Direct acting antiviral(DAA)treatments may reduce the elevatedαfetoprotein(AFP),but data on how these treatments affect elevated AFP in patients with chronic hepatitis C(CHC)remain insufficient.In the present study,the frequency of baseline AFP elevations and their related factors,AFP dynamics during and after DAA treatment,and factors associated with AFP reduction was assessed.This retrospective study included 141 patients with CHC without hepatocellular carcinoma who received DAA and achieved sustained virological response.The details are as follows:mean post-treatment follow-up was 99 weeks(12–213);mean age,57.8 years old;52%,males;79%,genotype(GT)1;and 47%,cirrhosis.Pre-treatment AFP elevation(>5.5 ng/mL)was seen in 48.2%patients.On multivariate analysis,baseline AFP>5.5 was associated with the presence of cirrhosis(P=0.001),co-existing non-alcoholic steatohepatitis(NASH)(P=0.035),and GT 1(P=0.029).AFP normalization was seen in 28.2%patients at treatment week 2,in 52%at the end of treatment,and in 73.4%at the end of follow-up.Post-treatment week 24 AFP normalization was associated with the absence of cirrhosis(P=0.003),Child--Pugh score<6(P=0.015),and baseline AFP<10(P=0.015).AFP elevation is common in patients with CHC and independently associated with NASH,cirrhosis,and GT 1.DAA treatment resulted in AFP normalization as early as treatment week 2.Post-treatment week 24 AFP normalization is independently associated with the absence of cirrhosis,Child--Pugh score<6,and baseline AFP<10.
基金This work is supported by the Foundationfor the Ninth F ive-Year Plan of PLA( 98JD3 2 69) Science and Tech-nology Development Fundation of Shanghai Municipality( 98XD14 0 2 0 ) "Hundred Leading Scientists Program" of the PublicHealth Sector
文摘To clone the murine α fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1 6 cells, and then the murine α fetoprotein gene was amplified by RT PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. After transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8kb murine α fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.
文摘Hepatocellular carcinoma(HCC) is the fifth most common cancer and the second leading cause of cancer-related deaths worldwide. Although the prognosis of patients with HCC is generally poor, the5-year survival rate is > 70% if patients are diagnosed at an early stage. However, early diagnosis of HCC is complicated by the coexistence of inflammation and cirrhosis. Thus, novel biomarkers for the early diagnosis of HCC are required. Currently, the diagnosis of HCC without pathological correlation is achieved by analyzing serum α.fetoprotein levels combined with imaging techniques. Advances in genomics and proteomics platforms and biomarker assay techniques over the last decade have resulted in the identification of numerous novel biomarkers and have improved the diagnosis of HCC. The most promising biomarkers,such as glypican-3, osteopontin, Golgi protein-73 and nucleic acids including microRNAs, are most likely to become clinically validated in the near future. These biomarkers are not only useful for early diagnosis of HCC, but also provide insight into the mechanisms driving oncogenesis. In addition, such molecular insight creates the basis for the development of potentially more effective treatment strategies. In this article,we provide an overview of the biomarkers that are currently used for the early diagnosis of HCC.
基金Supported by The Industrial Core Technology Development Program funded by the Ministry of Knowledge Economy,No.10033183
文摘AIM:To evaluate diagnostic value ofα-fetoprotein (AFP)-L3 and prothrombin induced by vitamin K absence-Ⅱ(PIVKA-Ⅱ)in hepatocellular carcinoma(HCC). METHODS:One hundred and sixty-eight patients during routine HCC surveillance were included in this study.Of the 168 patients,90(53.6%)had HCC including newly developed HCC(n=82)or recurrent HCC after treatment(n=8).Sera were obtained during their first evaluation for HCC development and at the time of HCC diagnosis before commencing HCC treatment.HCC was diagnosed by histological examination,appropriate imaging characteristics-computed tomography or magnetic resonance imaging.Control sera were collected from 78 patients with benign liver disease(BLD),which were obtained during routine surveillance with a suspicion of HCC.AFP,AFP-L3 and PIVKA-Ⅱwere measured in the same serum by microchip capillary electrophoresis and liquid-phase binding assay on a micro-total analysis system Wako i30 auto analyzer.The performance characteristics of three tests and combined tests for the diagnosis of HCC were obtained using receiver operating characteristic curves in all populations and subgroups with AFP<20 ng/mL. RESULTS:Of 90 HCC patients,38(42.2%)patients had AFP<20 ng/mL,20(22.2%)patients had AFP 20-200 ng/mL and 32(35.6%)patients had AFP>200 ng/mL.Of the 78 BLD patients,74(94.9%)patients had AFP<20 ng/mL.After adjustment for age and HBV infection status,AFP-L3 levels were higher in HCC than in BLD among patients with low AFP levels(<20 ng/mL)(P<0.001).In a total of 168 patients,areas under the curve(AUC)for HCC were 0.879,0.887,0.801 and 0.939 for AFP,AFP-L3,PIVKA-Ⅱand the combined markers,respectively.The combined AUC for three markers showed higher value than the AUCs of individual marker(P<0.05).AFP-L3 had higher AUC value than PIVKA-Ⅱfor HCC detection in entire patients(P =0.043).With combination of AFP-L3(cut-off>5%) and PIVKA-Ⅱ(cut-off>40 AU/L),the sensitivity were 94.4%and specificity were 75.6%in all patients.In 112 patients with low AFP levels(<20 ng/mL),AUCs
基金Supported by State Key Project Specialized for Infectious Diseases, No 2008ZX10002-015Beijing Municipal Health Bureau Key Project for Capital Medical Development, No 2007-1021
文摘AIM:To evaluate the diagnostic value of glypican-3(GPC3) in serum and liver for primary hepatocellular carcinoma(HCC).METHODS:Serum levels of GPC3 and α-fetoprotein(AFP) were measured in 75 patients with primary HCC and 32 patients with liver cirrhosis.Expression of GPC3 and AFP in 58 HCC and 12 cirrhotic specimens was detected with immunohistochemical staining.RESULTS:When the cut-off value of serum GPC3 was set at 300 ng/L,its sensitivity and specificity for HCC were 47.0% and 93.5%,respectively.Among the 14 patients with HCC at stage according to the Barcelona Clinic Liver Cancer staging system,the serum GPC3 level was higher than 300 ng/L in 50%(7/14) patients,the serum AFP level was not ≥ 400 μg/L in any patient.Combined serum AFP and GPC3 significantly increased the sensitivity to the diagnosis of HCC.The GPC3 expression was detected in cytoplasm of HCC cells but not in hepatocytes and bile ducts of benign tumors.Among the 58 HCC patients,the GPC3 was expressed in 100%(28/28) patients with their serum AFP level ≥ 400 μg/L,and in 90%(27/30) patients with their AFP level < 400 μg/L,respectively.The GPC3 was weakly or negatively expressed in all paracarcinomatous and cirrhotic tissue samples.AFP positive HCC cells were only found in 1 out of the 58 HCC patients.CONCLUSION:GPC3 protein is a sensitive and specific serum marker for diagnosis of early HCC.Its expression in liver tissues can be used to discriminate tumor cells from benign hepatic cells.
