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首页> 外文期刊>International journal of molecular medicine >Evaluation of annexin II as a potential serum marker for hepatocellular carcinoma using a developed sandwich ELISA method.
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Evaluation of annexin II as a potential serum marker for hepatocellular carcinoma using a developed sandwich ELISA method.

机译：使用开发的夹心ELISA方法评估Annexin II作为肝细胞癌的潜在血清标志物。

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Annexin II (Annexin A2, ANXA2) is a 36 kDa calcium-dependent phospholipid-binding protein that is located on the surface of most eukaryotic cells. ANXA2 is involved in several biological processes, including anti-inflammatory effects, Ca27+-dependent exocytosis, immune responses, Ca2+ transport and phospholipase A2 regulation. In our previous study, ANXA2 was identified as an up-regulated gene in hepatocellular carcinoma (HCC) tissue by cDNA microarray. In the present study, we have evaluated ANXA2 as a tumor-associated marker of HCC. We determined the ANXA2 levels in human liver tissues with HCC using real-time RT-PCR and Western blot analysis. For quantitative analysis of the ANXA2 protein in body fluids, we developed a sandwich ELISA system in which a polyclonal antibody and a monoclonal antibody specific to ANXA2 were employed as a capture antibody and a probe antibody, respectively. We detected the ANXA2 concentration in human serum using our newly developed system and evaluated its usefulness as a tumor marker. Overexpression of ANXA2 in human liver tissue was confirmed by real-time RT-PCR and Western blot analysis. The sandwich ELISA system for ANXA2 was developed for the detection of ANXA2 in human samples. The dose-response relationship between ANXA2 and optical density was linear in the range of 0-10 microg/ml and the sensitivity was 0.02 microg/ml. We determined the ANXA2 concentration in serum specimens using the newly developed sandwich ELISA. The serum ANXA2 concentrations of the patients with HCC (53.38+/-36.23 microg/ml) were significantly elevated when compared with those of normal individuals (28.81+/-24.94 microg/ml). These results suggest that expression of ANXA2 may be increased in HCC patients and may play an important role in liver cancer progression. This new ELISA method can be used as a tool for the detection of ANXA2 in human serum, particularly for cancer diagnostics.
机译：膜联蛋白II（膜联蛋白A2，ANXA2）是一种36 kDa的钙依赖性磷脂结合蛋白，位于大多数真核细胞的表面。 ANXA2参与多个生物学过程，包括抗炎作用，Ca27 +依赖的胞吐作用，免疫应答，Ca2 +转运和磷脂酶A2调节。在我们先前的研究中，ANXA2被cDNA微阵列鉴定为肝细胞癌（HCC）组织中的上调基因。在本研究中，我们评估了ANXA2作为HCC的肿瘤相关标志物。我们使用实时逆转录-聚合酶链反应和蛋白质印迹分析确定了肝癌患者肝组织中的ANXA2水平。为了定量分析体液中的ANXA2蛋白，我们开发了一种夹心ELISA系统，其中将对ANXA2特异的多克隆抗体和单克隆抗体分别用作捕获抗体和探针抗体。我们使用新开发的系统检测了人血清中ANXA2的浓度，并评估了其作为肿瘤标志物的有效性。实时RT-PCR和Western印迹分析证实了人肝组织中ANXA2的过度表达。开发了用于ANXA2的三明治ELISA系统，用于检测人类样品中的ANXA2。 ANXA2与光密度之间的剂量反应关系在0-10微克/毫升的范围内呈线性关系，灵敏度为0.02微克/毫升。我们使用最新开发的夹心ELISA测定了血清标本中的ANXA2浓度。与正常人（28.81 +/- 24.94 microg / ml）相比，HCC患者的血清ANXA2浓度（53.38 +/- 36.23 microg / ml）显着升高。这些结果表明，ANXA2的表达可能在肝癌患者中增加，并且可能在肝癌的进展中起重要作用。这种新的ELISA方法可用作检测人血清中ANXA2的工具，尤其是用于癌症诊断。

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