Molecular Pathology Unit, Cancer Research Centre, Institute for Medical Research (IMR), Kuala Lumpur, Malaysia.
Pathology. 2009;41(6):561-5. doi: 10.1080/00313020903071504.
Nasopharyngeal carcinoma (NPC) is a common malignancy among men in Malaysia. To determine the role of p53 in NPC, we screened for p53 mutations and evaluated the protein expression levels in samples from local patients with NPC.
Fifty-three formalin-fixed, paraffin-embedded nasopharyngeal carcinoma tissue blocks were chosen for this study. The presence of Epstein-Barr virus (EBV) was determined by in situ hybridisation using an EBER probe. p53 protein expression was detected using immunohistochemistry. Simultaneously, amplifications by PCR were performed for p53 exons 5 to 8, followed by mutation screening via single strand conformation polymorphism (SSCP). Sequencing of all the four exons was performed in five samples with mobility shift. To rule out false negative results by SSCP, 13 samples with p53 overexpression and five samples with low p53 expression were randomly selected and sequenced.
There was no mutation found in exons 5 to 8 in all the samples despite 46 (87%) of them having high p53 levels. EBV was detected in 51 (96%) out of 53 samples. There was no statistically significant association between p53 expression level and EBV presence.
High-intensity staining for p53 by immunohistochemistry was common in our series of NPC tissue samples but was not associated with 'hot spot' mutations of exons 5-8 of the gene. We did not find a significant relationship between the expression level of p53 and presence of EBV. Our study confirms that mutation of the DNA-binding domain of p53 is rare in NPC.
鼻咽癌(NPC)是马来西亚男性中常见的恶性肿瘤。为了确定 p53 在 NPC 中的作用,我们筛选了 p53 突变,并评估了来自当地 NPC 患者样本中的蛋白表达水平。
选择了 53 例福尔马林固定、石蜡包埋的鼻咽癌组织块进行本研究。使用 EBER 探针通过原位杂交确定 Epstein-Barr 病毒(EBV)的存在。使用免疫组织化学检测 p53 蛋白表达。同时,通过 PCR 进行 p53 外显子 5 到 8 的扩增,随后通过单链构象多态性(SSCP)进行突变筛选。对具有迁移变化的五个样本进行所有四个外显子的测序。为了排除 SSCP 的假阴性结果,随机选择了 13 个 p53 过表达和 5 个低 p53 表达的样本进行测序。
尽管 46 例(87%)样本中 p53 水平较高,但在所有样本中均未发现外显子 5 到 8 中的突变。在 53 例样本中,有 51 例(96%)检测到 EBV。p53 表达水平与 EBV 存在之间无统计学显著关联。
我们系列的 NPC 组织样本中,p53 的免疫组织化学高强度染色很常见,但与基因外显子 5-8 的“热点”突变无关。我们未发现 p53 表达水平与 EBV 存在之间存在显著关系。我们的研究证实 p53 的 DNA 结合域突变在 NPC 中很少见。