Assessment of apoptosis by immunohistochemical markers compared to cellular morphology in ex vivo-stressed colonic mucosa
Assessment of apoptosis by immunohistochemical markers compared to cellular morphology in ex vivo-stressed colonic mucosa
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DOI：
10.1369/jhc.4a6386.2005
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发表时间：
2005-02-01
影响因子：
3.2
通讯作者：
Garewal, H
中科院分区：
文献类型：
--
作者：
Holubec, H;Payne, CM;Garewal, H
细胞凋亡能力是预防癌症的核心。在结肠组织样品受到应激后，凋亡细胞的频率可用于测量凋亡能力，从而测量对结肠癌的可能易感性。评估细胞凋亡的金标准是形态学评价，但这需要有经验的显微镜。适用于存档石蜡包埋组织的细胞凋亡免疫组织化学标记物已在商业上开发。潜在有用的凋亡标志物包括裂解的细胞角蛋白-18（c-CK 18）、裂解的半胱天冬酶-3（c-cas-3）、裂解的核纤层蛋白A（c-lam-A）、磷酸化组蛋白H2 AX（gammaH 2AX）、裂解的聚（ADP核糖）聚合酶（c-PARP）和凋亡诱导因子（AIF）的易位。当从新鲜切除的结肠段的组织样本进行挑战离体与胆汁酸脱氧胆酸盐，类似于50%的杯状细胞成为凋亡的形态标准。这种高水平的形态学细胞凋亡允许与细胞凋亡免疫组织化学标记物的有用性和特异性进行定量比较。抗c-CK 18抗体对于鉴定凋亡结肠上皮细胞几乎与形态学一样有用和特异。针对c-cas-3、c-lam-A和gammaH 2AX的抗体虽然对凋亡细胞具有特异性，但用处不大。在我们的条件下，c-PARP的抗体虽然对凋亡细胞具有特异性，但有用性较低，而AIF的抗体相对非特异性。
Apoptosis competence is central to the prevention of cancer. Frequency of apoptotic cells, after a sample of colonic tissue is stressed, can be used to gauge apoptosis competence and, thus, possible susceptibility to colon cancer. The gold standard for assessment of apoptosis is morphological evaluation, but this requires an experienced microscopist. Easier-to-use immunohistochemical markers of apoptosis, applicable in archived paraffin-embedded tissue, have been commercially developed. Potentially useful apoptosis markers include cleaved cytokeratin-18 (c-CK18), cleaved caspase-3 (c-cas-3), cleaved lamin A (c-lam-A), phosphorylated histone H2AX (gammaH2AX), cleaved poly(ADP ribose) polymerase (c-PARP), and translocation of apoptosis-inclucing factor (AIF). When tissue samples from freshly resected colon segments were challenged ex vivo with the bile acid deoxycholate, similar to50% of goblet cells became apoptotic by morphologic criteria. This high level of morphologic apoptosis allowed quantitative comparison with the usefulness and specificity of immunohistochemical markers of apoptosis. The antibody to c-CK18 was almost as useful and about as specific as morphology for identifying apoptotic colonic epithelial cells. Antibodies to c-cas-3, c-lam-A, and gammaH2AX, though specific for apoptotic cells, were less useful. The antibody to c-PARP, though specific for apoptotic cells, had low usefulness, and the antibody to AIF was relatively nonspecific, under our conditions.