Application of a yeast assay to detect functional p53 mutations in archival prostate cancer tissue
Application of a yeast assay to detect functional p53 mutations in archival prostate cancer tissue
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DOI：
10.1089/108497802320970262
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发表时间：
2002-12-01
影响因子：
3.4
通讯作者：
White, RWD
中科院分区：
文献类型：
--
作者：
Shi, XB;Di Mauro, SM;White, RWD
使用酵母试验检测和功能评价突变型p53等位基因可以产生预测前列腺癌（CaP）患者预后的重要信息。由于该酵母菌检测试剂盒的当前版本不适用于存档组织，因此我们开发了一种用于福尔马林固定、石蜡包埋组织的改良检测试剂盒，并将其应用于患者样本的研究。使用这种改进的检测方法，我们检查了档案CaP样品从10例p53基因外显子5-8的突变。在四个样品中检测到突变：三个导致红色酵母菌落的形成，表明功能完全丧失，而一个产生粉红色酵母菌落，表明该突变体保留了部分功能。同时，我们使用单链构象多态性（SSCP）方法分析这些样本的p53异常。4份酵母菌阳性样品中只有3份出现异常SSCP条带。在两种方法均发现异常p53的情况下，DNA测序显示相同的碱基变化。这些结果表明，改进的酵母试验可能比SSCP检测p53突变更敏感，并证明改进的方法可用于检测和评价档案组织中存在的p53突变体的功能。
Detection and functional evaluation of mutant p53 alleles using a yeast assay could yield significant information for predicting the prognosis of patients with prostate cancer (CaP). Since the current version of this yeast assay is not applicable to archival tissues, we developed a modified assay for use on formalin-fixed, paraffin-embedded tissue and have applied it to the study of patient samples. Using this modified assay, we examined archival CaP samples from 10 patients for mutations in exons 5-8 of p53 gene. Mutations were detected in four samples: three resulted in the formation of red yeast colonies indicating complete loss of function, while one gave pink yeast colonies, indicating that this mutant retained partial function. In parallel, we analyzed these samples for p53 abnormalities using a single-strand conformational polymorphism (SSCP) approach. Only three of the four yeast-positive samples gave abnormal SSCP bands. In each case where abnormal p53 was found by both methods, DNA sequencing revealed the identical base change. These results suggest that the modified yeast assay may be more sensitive than SSCP for detection of p53 mutations, and demonstrate that the modified method can be used to detect and evaluate the function of p53 mutants present in archival tissue.