A Robust Immunohistochemical Assay for Detecting PTEN Expression in Human Tumors
A Robust Immunohistochemical Assay for Detecting PTEN Expression in Human Tumors
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DOI：
10.1097/pai.0b013e3181f1da13
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发表时间：
2011-03-01
影响因子：
1.6
通讯作者：
Stone, Steven
中科院分区：
文献类型：
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作者：
Sangale, Zaina;Prass, Cynthia;Stone, Steven
10 号染色体上缺失的磷酸酶和张力蛋白同源物 (PTEN) 是磷酸肌醇 3 激酶 (PI3K)/AKT 信号通路的负调节因子，控制细胞周期进程、生长和抑制细胞凋亡。 PTEN 蛋白表达缺失与肿瘤发生、癌症进展和耐药性相关，但存在相互矛盾的结果，部分原因可能是 PTEN 免疫组织化学 (IHC) 固有的困难。我们寻求一种稳健的 PTEN IHC 检测方法。通过拷贝数分析验证具有 PTEN 状态的人肿瘤细胞系进行福尔马林固定并石蜡包埋，用作阳性和阴性对照。 PTEN 抗体针对肿瘤细胞系进行了优化。对 10 个分子特征子宫内膜癌样本分析了 5 种优化抗体。四种抗体（CST、Millipore、Abcam、Novus）染色为 3/10 阳性和 7/10 阴性，然而，除 CST 之外的所有抗体都表现出阴性对照的非特异性核仁染色。一种抗体 (Dako) 染色为 5/10 阳性和 5/10 阴性，但区域为 (
Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a negative regulator of the phosphoinositol-3- kinase (PI3K)/AKT signaling pathway that controls cell cycle progression, growth and inhibition of apoptosis. Loss of PTEN protein expression has been associated with tumorigenesis, cancer progression and drug resistance, but conflicting results exist which may be due in part to difficulties inherent in PTEN immunohistochemistry (IHC). We sought a robust PTEN IHC assay. Human tumor cell lines with PTEN status verified by copy number analysis were formalin fixed and paraffin embedded for use as positive and negative controls. PTEN antibodies were optimized on tumor cell lines. Five optimized antibodies were analyzed on 10 molecularly characterized endometrial carcinoma samples. Four antibodies (CST, Millipore, Abcam, Novus) stained 3/10 positive and 7/10 negative, however, all but CST exhibited nonspecific nucleolar staining of negative controls. One antibody (Dako) stained 5/10 positive and 5/10 negative but with areas (