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目的：采用多重ＰＣＲ法检测喉癌及癌旁组织中Ｐ１６ 基因纯合缺失和异常甲基化。方法：选用二对引物分别扩增Ｐ１６ 基因外显子１ 和２，分析有无纯合缺失，对未检出缺失的样品用甲基化敏感内切酶Ｓｍ ａＩ消化后，再扩增外显子１，分析有无Ｐ１６ 基因异常甲基化。结果：３９ 例喉癌组织标本中９ 例标本有Ｐ１６ 基因纯合缺失，缺失率为２３．０８％ （９／３９）。３０ 例未检测到Ｐ１６ 基因缺失的喉癌组织标本，有４ 例检出异常甲基化，检出率为１３．３３％ （４／３０）。３９ 例癌旁组织标本均未检测到Ｐ１６基因缺失或异常甲基化。结论：采用多重ＰＣＲ法可检测出Ｐ１６ 基因常见的纯合缺失和异常甲基化失活。研究表明，Ｐ１６ 基因失活可能在喉癌的发生发展中起重要作用
Objective: To detect homozygous deletion and abnormal methylation of P16 gene in laryngeal squamous cell carcinoma and adjacent tissues by multiplex PCR. METHODS: Two pairs of primers were used to amplify the exon 1 and 2 of the P16 gene, respectively, to analyze the presence or absence of homozygous deletions. After the samples without deletions were digested with the methylation-sensitive endonuclease Sm aI, they were amplified. Sub-1 analysis of abnormal methylation of P16 gene. RESULTS: The homozygous deletion of P16 gene was detected in 9 specimens from 39 cases of laryngeal carcinoma. The missing rate was 23.08% (9/39). Thirty cases of laryngeal carcinoma without P16 gene deletion detected abnormal methylation in 4 cases, and the detection rate was 13.33% (4/30). The absence of P16 gene deletion or abnormal methylation was not detected in 39 paracancerous tissues. Conclusion: The homozygous deletion and abnormal methylation inactivation of the P16 gene can be detected by multiplex PCR. Studies show that P16 gene inactivation may play an important role in the development of laryngeal cancer