BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. We analyzed the expression of miclear-transcription factor-kappa B (NF-kappa B) during hepatocarcinogenesis in order to evaluate i...BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. We analyzed the expression of miclear-transcription factor-kappa B (NF-kappa B) during hepatocarcinogenesis in order to evaluate its dynamic expression and its clinical value in the development and diagnosis of HCC. METHODS: Hepatoma models were induced by oral administration of 2-acetamidoflurene (2-FAA) to male Sprague-Dawley rats. Morphological changes were observed after hematoxylin and eosin staining. The cellular distribution of NF-kappa B expression during different stages of cancer development was investigated by immunohistochemistry, and the level of NF-kappa B expression in liver tissues was quantitatively analyzed by ELISA. The gene fragments of hepatic NF-kappa B were amplified by nested-polymerase chain reaction assay. RESULTS: Hepatocytes showed vacuole-like degeneration during the early stages, then had a hyperplastic nodal appearance during the middle stages, and finally progressed to tubercles of cancerous nests with high differentiation. The NF-kappa B-positive material was buff-colored, fine particles localized in the nucleus, and the incidence of NF-kappa B-positive cells was 81.8% in degeneration, 83.3% in precancerous lesions, and 100% in cancerous tissues. All of these values were higher than those in controls (P<0.01). Hepatic NF-kappa B expression and hepatic NF-kappa B-mRNA were also higher during the course of HCC development (P<0.01). CONCLUSION: The NF-kappa B signal transduction pathway is activated during the early stages of HCC development, and its abnormal expression may be associated with the occurrence of HCC.展开更多
载脂蛋白B m RNA编辑酶催化多肽3F(APOBEC3F)属固有免疫系统中的重要成员,对人免疫缺陷病毒(HIV)、乙型肝炎病毒(HBV)等多种病毒的复制具有广泛的抑制作用。为构建APOBEC3F基因的真核表达载体,并检测其在体外培养的MARC145细胞中的表达...载脂蛋白B m RNA编辑酶催化多肽3F(APOBEC3F)属固有免疫系统中的重要成员,对人免疫缺陷病毒(HIV)、乙型肝炎病毒(HBV)等多种病毒的复制具有广泛的抑制作用。为构建APOBEC3F基因的真核表达载体,并检测其在体外培养的MARC145细胞中的表达情况,通过RT-PCR从猪脾脏组织扩增APOBEC3F基因,定向克隆到表达增强型绿色荧光蛋白(EGFP)的真核表达载体p EGFP-CMV中,构建p EGFP-APOBEC3F。PCR扩增及测序显示,APOBEC3F插入载体位置、方向及序列均正确。p EGFP-APOBEC3F经脂质体介导法转染MARC145细胞,转染后24 h APOBEC3F m RNA的水平升至最高。细胞免疫化学法检测发现APOBEC3F蛋白主要在MARC145细胞质中表达。研究结果为体外研究猪APOBEC3F基因在抗猪蓝耳病中的作用奠定了基础。展开更多
基金supported by grants from the Project of Elitist Peak in Six Fields(No.2006-B-063)the Projectof Medical Sciences(H200727),the Bureau of Health,Jiangsu Province,China
文摘BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. We analyzed the expression of miclear-transcription factor-kappa B (NF-kappa B) during hepatocarcinogenesis in order to evaluate its dynamic expression and its clinical value in the development and diagnosis of HCC. METHODS: Hepatoma models were induced by oral administration of 2-acetamidoflurene (2-FAA) to male Sprague-Dawley rats. Morphological changes were observed after hematoxylin and eosin staining. The cellular distribution of NF-kappa B expression during different stages of cancer development was investigated by immunohistochemistry, and the level of NF-kappa B expression in liver tissues was quantitatively analyzed by ELISA. The gene fragments of hepatic NF-kappa B were amplified by nested-polymerase chain reaction assay. RESULTS: Hepatocytes showed vacuole-like degeneration during the early stages, then had a hyperplastic nodal appearance during the middle stages, and finally progressed to tubercles of cancerous nests with high differentiation. The NF-kappa B-positive material was buff-colored, fine particles localized in the nucleus, and the incidence of NF-kappa B-positive cells was 81.8% in degeneration, 83.3% in precancerous lesions, and 100% in cancerous tissues. All of these values were higher than those in controls (P<0.01). Hepatic NF-kappa B expression and hepatic NF-kappa B-mRNA were also higher during the course of HCC development (P<0.01). CONCLUSION: The NF-kappa B signal transduction pathway is activated during the early stages of HCC development, and its abnormal expression may be associated with the occurrence of HCC.
文摘载脂蛋白B m RNA编辑酶催化多肽3F(APOBEC3F)属固有免疫系统中的重要成员,对人免疫缺陷病毒(HIV)、乙型肝炎病毒(HBV)等多种病毒的复制具有广泛的抑制作用。为构建APOBEC3F基因的真核表达载体,并检测其在体外培养的MARC145细胞中的表达情况,通过RT-PCR从猪脾脏组织扩增APOBEC3F基因,定向克隆到表达增强型绿色荧光蛋白(EGFP)的真核表达载体p EGFP-CMV中,构建p EGFP-APOBEC3F。PCR扩增及测序显示,APOBEC3F插入载体位置、方向及序列均正确。p EGFP-APOBEC3F经脂质体介导法转染MARC145细胞,转染后24 h APOBEC3F m RNA的水平升至最高。细胞免疫化学法检测发现APOBEC3F蛋白主要在MARC145细胞质中表达。研究结果为体外研究猪APOBEC3F基因在抗猪蓝耳病中的作用奠定了基础。
