Up-regulation of ALG-2 cancer tissue in hepatomas and lung
Up-regulation of ALG-2 cancer tissue in hepatomas and lung
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DOI：
10.1016/s0002-9440(10)63632-2
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发表时间：
2003-07-01
影响因子：
6
通讯作者：
Berchtold, MW
中科院分区：
文献类型：
--
作者：
la Cour, JM;Mollerup, J;Berchtold, MW
ALG - 2是在对参与程序性细胞死亡的蛋白质进行筛选时被分离出来的，并且是首个被发现直接参与细胞凋亡的钙离子结合蛋白。我们已经制备了适用于通过不同免疫学方法检测ALG - 2的多克隆抗体。三种商业化的抗ALG - 2抗体既不能检测小鼠重组ALG - 2，也不能检测Jurkat细胞裂解物中的内源性ALG - 2，然而我们自己亲和纯化的抗体能够识别重组的以及内源性的ALG - 2。通过预吸收实验以及在ALG - 2缺陷细胞上利用蛋白质印迹分析和免疫组织化学方法证明了该抗体的特异性。对15种不同的成年小鼠组织进行的蛋白质印迹分析表明，ALG - 2普遍表达。我们通过蛋白质印迹分析发现，与正常大鼠肝脏相比，大鼠肝癌中ALG - 2的表达量高出三倍多，免疫组织化学分析也证实了这一结果。对四种不同的肺癌组织微阵列（包括263名患者的样本）进行染色显示，ALG - 2主要定位于上皮细胞，并且在小细胞肺癌和非小细胞肺癌中显著上调。我们的研究结果得出这样的结论：除了已知的促凋亡功能外，ALG - 2可能在存活通路中也起作用。
ALG-2 was isolated in a screen for proteins involved in programmed cell death and is the first Ca2+-binding protein found to be directly involved in apoptosis. We have generated polyclonal antibodies that are suitable for detecting ALG-2 using different immunological methods. Three commercial antibodies against ALG-2 did neither detect mouse recombinant ALG-2 nor endogenous ALG-2 in Jurkat cell lysates, whereas our own affinity-purified antibody recognized recombinant as well as endogenous ALG-2. The specificity of the antibody was shown by preabsorbtion experiments and on ALG-2-deficient cells using Western blot analysis and immunohistochemistry. Western blot analysis of 15 different adult mouse tissues demonstrated that ALG-2 is ubiquitously expressed. We found that ALG-2 was more than threefold overexpressed in rat liver hepatoma compared to normal rat liver using Western blot analysis, a result confirmed by immunohistochemical analysis. Staining of four different lung cancer tissue microarrays; including specimens of 263 patients showed that ALG-2 is mainly localized to epithelial cells and significantly up-regulated in small-cell lung cancers and in nonsmall-cell lung cancers. Our results lead to the conclusion that ALG-2 beside its known proapoptotic functions may be a player in survival pathways.