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The purpose of the present study was to observe the expression of Axin protein during cardiac remodeling in rats. Cardiac remodeling animal models were prepared with the methods of jugular venous norepinephrine (NE)-infusion or arterial-vein fistula (AVF). The ultrasonic parameters of rat hearts were recorded before sacrifice. The expressions of Axin protein were determined by Western blot in rat hearts from different remodeling models as well as cultured cardiac fibroblasts from adult rats. Cardiac... The purpose of the present study was to observe the expression of Axin protein during cardiac remodeling in rats. Cardiac remodeling animal models were prepared with the methods of jugular venous norepinephrine (NE)-infusion or arterial-vein fistula (AVF). The ultrasonic parameters of rat hearts were recorded before sacrifice. The expressions of Axin protein were determined by Western blot in rat hearts from different remodeling models as well as cultured cardiac fibroblasts from adult rats. Cardiac concentric hypertrophy and fibrosis was induced by 3-day jugular vein infusion of NE in rats. The expression of Axin in the left ventricles increased significantly compared with that of the control group. Cardiac eccentric hypertrophy without fibrosis was induced by A-V fistula for one week in rats, and no change in Axin protein expression in the left ventricles was observed. In cultured adult rat cardiac fibroblasts, NE treatment for 24 h increased significantly the Axin protein level. It is therefore concluded that Axin protein was expressed in rat heart and increased significantly in left ventricles during NE-induced rat cardiac remodeling, which may be relevant to cardiac fibrosis. 为观察大鼠心肌重塑过程中Axin蛋白质表达水平的变化 ,实验用颈静脉输注去甲肾上腺素 (NE)和动静脉造瘘 (AVF)方法复制大鼠心肌重塑病理模型 ,采用超声心动术检测心脏结构和收缩功能。取病理模型大鼠左心室以及分离培养的成年大鼠心肌成纤维细胞 ,采用Westernblot技术检测Axin蛋白质的表达水平。结果观察到 ,在颈静脉输注NE 3d后 ,大鼠心脏发生向心性心肌肥厚和心肌纤维化 ,其左心室的Axin蛋白表达水平较对照组显著升高。A V造瘘术一周后引起大鼠离心性心肌肥厚 ,心肌无明显纤维化 ,心肌Axin表达量与对照相比无显著变化。在分离培养的成年大鼠心肌成纤维细胞 ,NE处理 2 4h能明显升高Axin蛋白的表达水平。上述结果表明 ,大鼠心脏有Axin蛋白质表达 ,NE致大鼠心肌重塑过程中Axin蛋白表达显著增加 ,可能与该过程的心肌纤维化有关 AIM: To construct the eukaryotic expression vector pIRES2-EGFP-Axin, and to express Axin in C6 glioma cells. METHODS: The Axin gene was amplified by PCR using pCMV5-HA-Axin as a template, and confirmed by DNA sequencing. The eukaryotic expression vector pIRES2-EGFP-Axin was constructed by introducing Axin DNA fragment into the sites of Nhe I and Sal I of pIRES2-EGFP vector. The plasmid was transfected into the C6 cells using lipofectamine. The expressed EGFP was observed under... AIM: To construct the eukaryotic expression vector pIRES2-EGFP-Axin, and to express Axin in C6 glioma cells. METHODS: The Axin gene was amplified by PCR using pCMV5-HA-Axin as a template, and confirmed by DNA sequencing. The eukaryotic expression vector pIRES2-EGFP-Axin was constructed by introducing Axin DNA fragment into the sites of Nhe I and Sal I of pIRES2-EGFP vector. The plasmid was transfected into the C6 cells using lipofectamine. The expressed EGFP was observed under fluorescent microscope and the Axin protein expression was detected by immunostaining using anti-Axin antibody. RESULTS: The eukaryotic expression vector pIRES2-EGFP-Axin was constructed and transfected successfully into C6 glioma cells. The green fluorescence of EGFP was observed in the plasma and nuclei of transfected cells, and Axin protein was only found in the plasma. CONCLUSION: The recombinant expression vector pIRES2-EGFP-Axin was constructed, and the EGFP and Axin gene could be co-expressed in the C6 cells. This study laid a foundation for the further research of the function of Axin in cell differentiation, growth and tumorigenesis. 目的:构建真核表达载体pIRES2EGFPAxin,并在神经胶质瘤细胞系C6中进行表达。方法:用PCR的方法扩增Axin基因,构建真核表达载体pIRES2EGFPAxin,经NheI及SalI双酶切鉴定并测序。通过脂质体法转染C6细胞,用荧光显微镜检测细胞中增强型绿色荧光蛋白(EGFP)的表达,用免疫组化染色的方法检测细胞中Axin的表达。结果:构建了真核表达载体pIRES2EGFPAxin,用脂质体法转染神经胶质瘤细胞C6后,经荧光显微镜和免疫组化染色法检测,可见细胞内有EGFP及Axin的表达。结论:成功地构建真核表达载体pIRES2EGFPAxin,并在神经胶质瘤细胞C6中表达,为研究Axin对肿瘤的生物学作用以及Axin在肿瘤基因治疗中的应用奠定了基础。 AIM: To investigate the relationship between Axin protein expression and genesis and metastasis of gastric carcinoma. METHODS: A total of 46 patients with gastric carcinoma underwent surgical resection were included in this study. The expression of Axin protein was detected by immunohistochemical techniques (SABC) in paraffin-embedded samples prepared from gastric carcinoma tissues and normal stomach tissues far from the tumor. In addition, the positive rate of Axin expression was calculated... AIM: To investigate the relationship between Axin protein expression and genesis and metastasis of gastric carcinoma. METHODS: A total of 46 patients with gastric carcinoma underwent surgical resection were included in this study. The expression of Axin protein was detected by immunohistochemical techniques (SABC) in paraffin-embedded samples prepared from gastric carcinoma tissues and normal stomach tissues far from the tumor. In addition, the positive rate of Axin expression was calculated and its corrrelation with the pathological characteristics of gastric carcinoma was analyzed. RESULTS: In normal stomach tissues, Axin pro tein expression was strongly positive and intensified in basal cells. Axin was also expressed in tissues from gastric carcinoma and normal stomach mucosa far from the tumor with a positive rate of 62.0% and 91.3%, respectively. There was a significant difference between them (P < 0.01). Axin protein expression was significantly correlated with clinical TNM classification (TNM Ⅰ, Ⅱ vs Ⅲ, Ⅳ: 78.6% vs 56.3%, P = 0.035) and lymphatic metastasis (without vs with: 85.7% vs 53.1%, P= 0.034). The expression of Axin protein was not markedly correlated with the age and gender of patients, tumor size, biological features and serosa invasion (P > 0.05). CONCLUSION: The expression of Axin protein is down-regulated in gastric carcinoma, which correlates with the genesis and metastasis of the disease. 目的:探讨胃癌组织Axin蛋白的表达与侵袭转移的相关性.方法:胃癌患者46例,均行胃癌根治切除手术.采集胃癌癌组织及远癌正常胃组织标本, 制备石蜡切片,采用免疫组化(SABC)法检测 Axin蛋白的表达,研究其表达和分布特点与临床病理间的关系.结果:正常胃组织Axin蛋白在基底部细胞表达强,表面成熟细胞中表达弱:胃癌组织、远癌正常胃组织中均有Axin蛋白表达,其阳性表达率分别为62.0%和91.3%,差异有统计学意义(P<0.01);Axin蛋白的表达与胃癌临床病理分期(TNM I,ⅡvsⅢ,Ⅳ:78.6% vs 56.3%, P=0.035)、有无淋巴转移有关(无 vs 有:85.7% vs 53.1%,P=0.034),与胃癌患者性别、年龄、肿瘤大小、生物学特征和是否侵及浆膜无关(P>0.05).结论:Axin蛋白表达减弱与胃癌的发生以及肿瘤的临床进展和淋巴转移相关.
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