He Guang, Fu Wei-Neng, Qiu Guang-Bin, Zhao Zhen, Xu Zhen-Ming, Sun Xing-He, Sun Kai-Lai
Department of Medical Genetics, China Medical University, Shenyang 110001, China.
Yi Chuan Xue Bao. 2002 May;29(5):390-5.
In order to investigate the role of Cytokeratin 13(CK13) gene in laryngeal carcinogenesis, we detected the deletion of CK13 gene through LOH analysis indirectly at DNA level using 5 STR primers within and near CK13 gene in 72 cases of laryngeal squamous cell carcinoma, then detected the differential expression between 16 cases of paired normal and cancerous tissue by Northern blot, and performed immunohistochemistry using well characterized monoclonal antibody against CK13 in squamous cell carcinoma of different stages. We found that all of the microsatellite loci exist LOH, and the LOH frequencies were 18.03%, 28.13%, 27.42%, 39.68% and 34.85% at D17S1964E, D17S2092, D17S791, D17S1665 and D17S808 respectively. The LOH+ cases accounted for 77.78% (56/72), and the frequencies of LOH were not related to the type of laryngeal carcinoma and the lymphoid metastasis; but significantly related to the differentiation, P < 0.05. CK13 gene is expressed significantly higher in 16 cases of normal tissues than in paired cancerous tissues, and the immunostain revealed that CK13 was expressed in normal laryngeal squamous cell or high differentiation stage, and its expression decreased or disappeared in poor ones, P < 0.01. CK13 gene might play an important role in the laryngeal carcinogenesis, acting as a novel tumor suppressor gene, and may be relevant to laryngeal squamous cell carcinoma diagnosis and prognosis. Further research will contribute to conform it.
为研究细胞角蛋白13(CK13)基因在喉癌发生中的作用,我们采用位于CK13基因内部及附近的5个STR引物,通过DNA水平的杂合性缺失(LOH)分析间接检测72例喉鳞状细胞癌中CK13基因的缺失情况;然后用Northern印迹法检测16例配对的正常组织和癌组织之间的差异表达;并使用针对CK13的特异性良好的单克隆抗体,对不同分期的鳞状细胞癌进行免疫组织化学检测。我们发现所有微卫星位点均存在LOH,在D17S1964E、D17S2092、D17S791、D17S1665和D17S808处的LOH频率分别为18.03%、28.13%、27.42%、39.68%和34.85%。LOH阳性病例占77.78%(56/72),LOH频率与喉癌类型及淋巴转移无关;但与分化程度显著相关,P<0.05。CK13基因在16例正常组织中的表达明显高于配对的癌组织,免疫染色显示CK13在正常喉鳞状细胞或高分化阶段表达,而在低分化者中表达降低或消失,P<0.01。CK13基因可能在喉癌发生中起重要作用,作为一种新的肿瘤抑制基因,可能与喉鳞状细胞癌的诊断和预后相关。进一步的研究将有助于证实这一点。