BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, its prognosis is poor, and early detection is of utmost importance. Although alpha-fetoprotein (AFP) is a useful marker ...BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, its prognosis is poor, and early detection is of utmost importance. Although alpha-fetoprotein (AFP) is a useful marker for detecting and monitoring HCC development, the false-negative or false-positive rates with AFP alone may be as high as 30%-40% for patients with small HCCs. To enhance the specificity and accuracy of AFP measurements for HCC, we analyzed AFP expression states in livers, detected the hepatoma-specific AFP (HS-AFP) fraction and AFP-mRNA from peripheral blood mononuclear cells, and explored their clinical implications for HCC diagnosis. METHODS: AFP expression and hepatocyte distributions in liver specimens were investigated by an immunohistoche- mical assay. Total RNAs were extracted from circulating blood, synthesized to cDNA through random primers and reverse transcriptase, and fragments of the AFP gene were amplified by a nested-PCR assay. The HS-AFP fraction was separated by lectin-affinity chromatography and its level was detected by radioimmunoassay. RESULTS: The incidence of AFP was 73.3% in HCC tissues and its expression in HCCs with moderate or low differentiation was significantly stronger than that of HCCs with high differentiation (P<0.05). The incidence of AFP gene fragments was 100% in HCCs, and 60% in paracancerous tissues (P<0.01). In the HCC and liver cirrhosis groups, the incidence of HS-AFP was 91.7% and 18% (P<0.01), and of AFP-mRNA was 56.7% and 16% (P<0.01), respectively, and neither was found in controls.HS-AFP or AFP-mRNA was not significantly related to size or number of HCC, but to its differentiation, metastasis, and relapse (P<0.05). CONCLUSIONS: Different AFP expression is present in different parts of HCC tissues. HS-AFP and AFP-mRNA fragments improve sensitivity and specificity, and both are useful markers to diagnose HCC or monitor metastasis and relapse.展开更多
Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was se...Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin （LCA）-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay （RIA）. Results： Circulating AFP was separated into three peaks （AFP-1, AFP-2, and AFP-3） by LCA-affinity chromatography. Dunng the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HSAFP （AFP-3） from sera of HCC patients was eluted clearly on the LCA-sepharose gel mini-column with a solution containing α-methyI-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity （92.7%） and specificity （88.2%）. Conclusion： The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.展开更多
Objective: Early finding of hepatocellular carcinoma(HCC) and monitoring of its metastasis are of the utmost importance. The objectives of this study were to investigate the values of circulating hepatoma-specific gam...Objective: Early finding of hepatocellular carcinoma(HCC) and monitoring of its metastasis are of the utmost importance. The objectives of this study were to investigate the values of circulating hepatoma-specific gamma-glutamyl transferase(HS-GGT) fraction and α-fetoprotein mRNA(AFP-mRNA) from peripheral blood mononuclear cells(PBMCs) in diagnosis or monitoring metastasis of HCC. Methods: Total RNA was extracted from hepatomas or PBMCs of patients, synthesized to AFP-cDNA through random primers and reverse transcriptase, amplified by using a nested PCR assay, and confirmed by sequencing. Simultaneity, the HS-GGT activities were quantitatively determined in sera of patients. The comprehensive assessments of two markers were investigated in HCC patients. Results: The amplified fragments of AFP-mRNA were identical to original designed ones(159 bp) and confirmed by sequencing. The lowest sensitivity was 2 ng/L of total RNA. The incidences of AFP-mRNA and HS-GGT were 60.4%(113 of 187) and 84.5%(158 of 187) in the HCC group, and their significantly higher(P < 0.01) than that in any of other groups. The high frequencies of HS-GGT and AFP-mRNA were 78.2% and 56.4% in the HCC patients with AFP < 50 ng/mL. All positive of AFP-mRNA was found in HCC patients with extrahepatic metastasis. No significant correlation was found between tumor size and AFP-mRNA or HS-GGT. The total incidence of combined AFP-mRNA and HS-GGT detection was 93.6 % for HCC diagnosis. Conclusion: HS-GGT and AFP-mRNA are specific markers for HCC diagnosis or differentiation, especially the former for early finding and the later for monitoring metastasis of HCC.展开更多
目的:探讨肝癌发生过程中肝癌特异性 ALP 的产生与变化。方法:以含有2-FAA(0.05%)饲料饲养 SD 大鼠制作肝癌模型,对癌变过程中,鼠血清和肝组织 ALP 和同功酶进行了分析。结果:病理学检查及生化学方法发现,诱癌后鼠肝大量表达出 ALP;并...目的:探讨肝癌发生过程中肝癌特异性 ALP 的产生与变化。方法:以含有2-FAA(0.05%)饲料饲养 SD 大鼠制作肝癌模型,对癌变过程中,鼠血清和肝组织 ALP 和同功酶进行了分析。结果:病理学检查及生化学方法发现,诱癌后鼠肝大量表达出 ALP;并引起血中 ALP 活性的异常:实验组血清总 ALP,肝组织总 ALP,可溶性及膜结合性 ALP 均高于对照组(P 值<0.01:);后两者的平均比,6周后各组显著升高;在聚丙烯酰胺凝胶垂直平板电泳上,ALP 同功酶可见5条区带:ALPⅠ和 ALP-Ⅱ分别为正常鼠肝和肝癌鼠肝所特有,6周时肝中出现 ALP-Ⅱ,第8周后肝及血中 ALP-Ⅱ同步出现。结论:鼠肝癌变过程中具有癌胚性质的 ALP 过度表达,异常 ALP-Ⅱ的分析有助于鼠肝癌的早期诊断。展开更多
基金This study was supported in part by a grant from the Project of the Bureau of Science and Technology, Nantong (s30033), China.
文摘BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, its prognosis is poor, and early detection is of utmost importance. Although alpha-fetoprotein (AFP) is a useful marker for detecting and monitoring HCC development, the false-negative or false-positive rates with AFP alone may be as high as 30%-40% for patients with small HCCs. To enhance the specificity and accuracy of AFP measurements for HCC, we analyzed AFP expression states in livers, detected the hepatoma-specific AFP (HS-AFP) fraction and AFP-mRNA from peripheral blood mononuclear cells, and explored their clinical implications for HCC diagnosis. METHODS: AFP expression and hepatocyte distributions in liver specimens were investigated by an immunohistoche- mical assay. Total RNAs were extracted from circulating blood, synthesized to cDNA through random primers and reverse transcriptase, and fragments of the AFP gene were amplified by a nested-PCR assay. The HS-AFP fraction was separated by lectin-affinity chromatography and its level was detected by radioimmunoassay. RESULTS: The incidence of AFP was 73.3% in HCC tissues and its expression in HCCs with moderate or low differentiation was significantly stronger than that of HCCs with high differentiation (P<0.05). The incidence of AFP gene fragments was 100% in HCCs, and 60% in paracancerous tissues (P<0.01). In the HCC and liver cirrhosis groups, the incidence of HS-AFP was 91.7% and 18% (P<0.01), and of AFP-mRNA was 56.7% and 16% (P<0.01), respectively, and neither was found in controls.HS-AFP or AFP-mRNA was not significantly related to size or number of HCC, but to its differentiation, metastasis, and relapse (P<0.05). CONCLUSIONS: Different AFP expression is present in different parts of HCC tissues. HS-AFP and AFP-mRNA fragments improve sensitivity and specificity, and both are useful markers to diagnose HCC or monitor metastasis and relapse.
基金grant from the Project of the Bureau of Science and Technology of Nantong (S30033).
文摘Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin （LCA）-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay （RIA）. Results： Circulating AFP was separated into three peaks （AFP-1, AFP-2, and AFP-3） by LCA-affinity chromatography. Dunng the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HSAFP （AFP-3） from sera of HCC patients was eluted clearly on the LCA-sepharose gel mini-column with a solution containing α-methyI-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity （92.7%） and specificity （88.2%）. Conclusion： The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.
基金Supported by grants from the National Natural Science Foundation of China(No.81200634)the Projects of Jiangsu Medical Science(No.WSW-011,HK201102,BL2012053)+1 种基金the Qinglan Project of Jiangsu higher Educationhe International S&T Cooperation Program of China(No.2013DFA32150)
文摘Objective: Early finding of hepatocellular carcinoma(HCC) and monitoring of its metastasis are of the utmost importance. The objectives of this study were to investigate the values of circulating hepatoma-specific gamma-glutamyl transferase(HS-GGT) fraction and α-fetoprotein mRNA(AFP-mRNA) from peripheral blood mononuclear cells(PBMCs) in diagnosis or monitoring metastasis of HCC. Methods: Total RNA was extracted from hepatomas or PBMCs of patients, synthesized to AFP-cDNA through random primers and reverse transcriptase, amplified by using a nested PCR assay, and confirmed by sequencing. Simultaneity, the HS-GGT activities were quantitatively determined in sera of patients. The comprehensive assessments of two markers were investigated in HCC patients. Results: The amplified fragments of AFP-mRNA were identical to original designed ones(159 bp) and confirmed by sequencing. The lowest sensitivity was 2 ng/L of total RNA. The incidences of AFP-mRNA and HS-GGT were 60.4%(113 of 187) and 84.5%(158 of 187) in the HCC group, and their significantly higher(P < 0.01) than that in any of other groups. The high frequencies of HS-GGT and AFP-mRNA were 78.2% and 56.4% in the HCC patients with AFP < 50 ng/mL. All positive of AFP-mRNA was found in HCC patients with extrahepatic metastasis. No significant correlation was found between tumor size and AFP-mRNA or HS-GGT. The total incidence of combined AFP-mRNA and HS-GGT detection was 93.6 % for HCC diagnosis. Conclusion: HS-GGT and AFP-mRNA are specific markers for HCC diagnosis or differentiation, especially the former for early finding and the later for monitoring metastasis of HCC.
文摘目的:探讨肝癌发生过程中肝癌特异性 ALP 的产生与变化。方法:以含有2-FAA(0.05%)饲料饲养 SD 大鼠制作肝癌模型,对癌变过程中,鼠血清和肝组织 ALP 和同功酶进行了分析。结果:病理学检查及生化学方法发现,诱癌后鼠肝大量表达出 ALP;并引起血中 ALP 活性的异常:实验组血清总 ALP,肝组织总 ALP,可溶性及膜结合性 ALP 均高于对照组(P 值<0.01:);后两者的平均比,6周后各组显著升高;在聚丙烯酰胺凝胶垂直平板电泳上,ALP 同功酶可见5条区带:ALPⅠ和 ALP-Ⅱ分别为正常鼠肝和肝癌鼠肝所特有,6周时肝中出现 ALP-Ⅱ,第8周后肝及血中 ALP-Ⅱ同步出现。结论:鼠肝癌变过程中具有癌胚性质的 ALP 过度表达,异常 ALP-Ⅱ的分析有助于鼠肝癌的早期诊断。
