用银染PCR-SSCP方法检测肺鳞癌组织标本中p16基因和E1.β外显子异常

Detection of alterations of p16 gene by silver staining PCR SSCP in lung squamous cell carcinoma tissue

目的：检测肺鳞癌组织中ｐ１６基因和Ｅ１．β外显子的异常改变，进一步观察该基因在肺鳞癌组织细胞周期调节紊乱中的异常，探讨肺鳞癌发生、发展的分子机制。方法：采用银染ＰＣＲ－ＳＳＣＰ方法对常规石蜡包埋肺鳞癌标本中ｐ１６基因和Ｅ１．β外显子异常进行检测。结果与结论：１１例免疫组化检测ｐ１６表达阴性的肺鳞癌标本中，４例有异常电泳带，其中，第１、２外显子各１例，第３外显子２例；从６例ｐ１６蛋白阳性肺癌组织检出１例Ｅ１．β外显子异常。提示ｐ１６基因和Ｅ１．β外显子突变或（和）缺失是肺鳞癌组织ｐ１６蛋白表达异常的可能原因之一。

Objective: To observe the alterations of p16 gene and E1. β in human lung squamous cell carcinomas and to study the molecular mechanism in the development of human lung squamous cell carcinomas. Methods: Silver staining PCR SSCP method was used to detect alterations of p16 gene and E1. β in 17 paraffin embedded human lung squamous cell carcinomas. Results and Conclusion: The abnormal shift of the single strand DNA(ssDNA) was identified in 4 out of 11 cases without detection of p16 protein by immunohistochemistry: two in exon 3, one in exon 1 and one in exon 2;one of 6 p16 positive tumors were found to be mutated in E1. β of p16. The above results suggested that mutation and/or deletion in p16 gene may be one of the causes of alterated expression of p16 protein and play a role in the development of human lung squamous cell carcinomas.