Liao P H, Chang Y C, Huang M F, Tai K W, Chou M Y
Department of Dentistry, Chung Shan Medical and Dental College Hospital, Taichung, Taiwan.
Oral Oncol. 2000 May;36(3):272-6. doi: 10.1016/s1368-8375(00)00005-1.
The inactivation of tumor suppressor gene (TSG) is important during multistage carcinogenesis. The p53 TSG is frequently mutated in oral squamous cell carcinomas. These mutations can serve as very specific markers for the presence of tumor cells in a background of normal cells. In this study, 10 oral squamous cell carcinoma patients and 27 normal dental students were collected from Chung Shan Medical and Dental College Hospital, Taichung, Taiwan. Extractions of DNA from saliva were obtained. Exon 4 and intron 6 within the p53 gene were amplified with polymerase chain reactions (PCRs) followed by DNA sequence analysis. DNA sequence analysis of PCR products revealed that five of eight (62.5%) tumor saliva samples and five of 27 (18. 52%) healthy saliva samples contained p53 exon 4 codon 63 mutations. These results were significantly different by using Chi-square test (P<0.05). The majority of the base substitutions were C deletions. Probable hot spots for the mutation were identified at exon 4 codon 63, which has not been observed before in head and neck cancers. Our study indicated that mutation of p53 codon 63 in saliva might be a molecular marker for oral squamous cell carcinomas. In addition, the amount of DNA recovered from saliva in most cases is sufficiently large and its quality suitable to enable PCR amplification which could be used in the search for mutations. The protocol described is rapid, cheap, and easy to perform, and may be useful for epidemiological studies for oral carcinogenesis.
肿瘤抑制基因(TSG)的失活在多阶段致癌过程中很重要。p53肿瘤抑制基因在口腔鳞状细胞癌中经常发生突变。这些突变可作为在正常细胞背景中存在肿瘤细胞的非常特异的标志物。在本研究中,从台湾台中市中山医学暨牙医学院附设医院收集了10例口腔鳞状细胞癌患者和27名正常牙科学生。从唾液中提取DNA。用聚合酶链反应(PCR)扩增p53基因的第4外显子和第6内含子,随后进行DNA序列分析。PCR产物的DNA序列分析显示,8份肿瘤唾液样本中的5份(62.5%)和27份健康唾液样本中的5份(18.52%)含有p53第4外显子密码子63突变。使用卡方检验,这些结果有显著差异(P<0.05)。大多数碱基替换是C缺失。在第4外显子密码子63处鉴定出可能的突变热点,这在头颈癌中以前未观察到。我们的研究表明,唾液中p53密码子63的突变可能是口腔鳞状细胞癌的分子标志物。此外,在大多数情况下,从唾液中回收的DNA量足够大,其质量适合进行PCR扩增,可用于寻找突变。所描述的方案快速、廉价且易于操作,可能对口腔癌发生的流行病学研究有用。