Lang J C, Tobin E J, Knobloch T J, Schuller D E, Bartynski K J, Mountain R E, Nicholson R, DeYoung B R, Weghorst C M
Department of Otolaryngology, Ohio State University Comprehensive Cancer Center, Arthur G. James Cancer Hospital and Research Institute, Columbus 43210, USA.
Laryngoscope. 1998 Jun;108(6):923-8. doi: 10.1097/00005537-199806000-00024.
RNA was isolated from 22 squamous cell carcinomas (SCCs) obtained from diverse sites within the head and neck and from matched normal tissue where available. Tissue samples were then screened for expression of RNA from tumor suppressor gene p16 by utilizing semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis. p16-Specific PCR amplification products generated from tumor samples were subject to further analysis by direct DNA sequencing to determine if any tumor sample harbored a p16 mutation. The results show the presence of mutations in 10 of 22 (45%) of the tumor samples. Mutations comprise two identical point mutations, two small deletions (1 bp and 2 bp), one single-nucleotide insertion, four larger deletions, and an insertion/deletion. No mutations in p16 have been identified by analysis of PCR products generated from normal matched tissue, suggesting that p16 alterations are generated by somatic mutation and are not germline in origin. All 22 samples were analyzed additionally by immunohistochemistry for nuclear expression of the retinoblastoma (RB) tumor suppressor gene product. Results show lack of RB nuclear expression in only one sample, suggesting that mutation of RB is an infrequent event in the development of SCC of the head and neck (SCCHN).
从22例头颈部不同部位的鳞状细胞癌（SCC）以及可获取的匹配正常组织中分离RNA。然后利用半定量逆转录聚合酶链反应（RT-PCR）分析筛选组织样本中肿瘤抑制基因p16的RNA表达。对肿瘤样本产生的p16特异性PCR扩增产物进行直接DNA测序进一步分析，以确定是否有任何肿瘤样本存在p16突变。结果显示，22个肿瘤样本中有10个（45%）存在突变。突变包括两个相同的点突变、两个小缺失（1 bp和2 bp）、一个单核苷酸插入、四个较大的缺失以及一个插入/缺失。通过对匹配正常组织产生的PCR产物分析，未发现p16突变，这表明p16改变是由体细胞突变产生的，并非种系起源。另外，对所有22个样本进行免疫组织化学分析，检测视网膜母细胞瘤（RB）肿瘤抑制基因产物的核表达。结果显示仅一个样本缺乏RB核表达，这表明RB突变在头颈部鳞状细胞癌（SCCHN）发生中是罕见事件。
