Pathological significance of oxidative stress and its application to pathologic diagnosis

硝酸三乙酸铁（Fe-NTA）诱导氧化性肾损伤导致大鼠肾细胞癌（RCC）的高发。这些rcc的差异显示分析显示膜联蛋白2 （Anx2）的表达升高，这是一种激酶的底物和组织型纤溶酶原的受体。我们进行这项研究是为了阐明Anx2在Fenton反应基础癌变中的意义。Fe-NTA处理后大鼠肾脏和H_2O_2处理后LLC-PK1细胞中Anx2信使RNA和蛋白水平呈时间依赖性升高。n -乙酰半胱氨酸、吡咯烷二硫代氨基甲酸酯或过氧化氢酶预处理对后者均有抑制作用。免疫组织化学显示正常肾近端小管染色可忽略，但再生肾近端小管、核细胞和肾细胞癌染色强烈。转移性rcc表现出较高的Anx2蛋白水平。在这些细胞中，Anx2在丝氨酸和酪氨酸残基处被磷酸化，并与磷酸化的肌动蛋白一起沉淀。在lc - pki细胞中，过表达Anx2可诱导更高的细胞增殖率。相反，对fe - nia诱导的rcc细胞系进行Anx2反义处理后，观察到增殖减少导致细胞凋亡。这些结果表明，Anx2受氧化还原状态的调控，这种适应性机制的持续运作在氧化应激诱导的癌症的增殖和转移中起着重要作用。此外，我们证明了基于肽核酸（PNA）的mRNA原位杂交技术适用于50年前生产的石蜡包埋病理标本。我们开发了一种极快速的方法，利用低剂量的微波辐射进行免疫印迹分析。最后，我们制定了abo血型不相容肝移植早期体液排斥的诊断标准。这对于研究氧化应激在移植肝排斥反应中的作用具有重要意义。少

Ferric nitrilotriacetate (Fe-NTA) induces oxidative renal damage leading to a high incidence of renal cell carcinoma (RCC) in rats. Differential display analysis of such RCCs revealed elevated expression of annexin 2 (Anx2), a substrate for kinases and a receptor for tissue-type plasminogen adivator and plasminogen. We conducted this study to clarifythe significance of Anx2 in Fenton reaction-based carcinogenesis. Messenger RNA and protein levels of Anx2 were increased time-dependently in the rat kidney after Fe-NTA administration as well as in LLC-PK1 cells after exposure to H_2O_2. The latter was inhibited by pretreatment with N-acetylcysteine, pyrrolidine dithiocarbamate or catalase. immunohistochemistry revealed negligible staining in the normal renal proximal tubules, but strong staining in regenerating proximal tubules, karyomegalic cells and RCCs. Metastasizing RCCs showed higher Anx2 protein levels. Anx2 was phosphorylated at serine and tyrosine residues in these cells and coim … More munoprecipitated with phosphorylated actin. Overexpresison of Anx2 induced a higher cell proliferation rate in LLC-PKI cells. In contrast, a decrease in proliferation leading to apoptosis was observed after Anx2 antisense treatment to cell lines established from Fe-NIA-induced RCCs. These results suggest that Anx2 is regulated by redox status, and that persistent operation of this adaptive mechanism plays a role in the proliferation and metastasis of oxidative stress-induced cancer. Furthermore, we demonstrated that peptide nucleic acid (PNA)-based mRNA in situ hybridization is applicable to paraffin-embedded pathologic specimens produced 50 years ago. We have developed an extremely rapid method for Western blot analysis by the use of low dose of microwave radiation. Finally, we have produced diagnostic criteria for the early humoral rejection in ABO-incompatible liver transplantation. This would be important for investigating the involvement of oxidative stress in the rejection of transplanted liver. Less