头颈鳞癌和颈清扫淋巴结D9S171位点杂合性缺失的研究

目的探讨头颈鳞状细胞癌在D9S171微卫星多态性位点的杂合性缺失(Loss of heterozygosity,LOH)的发生率、临床意义及对检测颈淋巴结转移的应用价值.方法标准酚氯仿法提取肿瘤组织及颈清扫淋巴结组织的基因组DNA,采用D9S171微卫星多态性位点进行聚合酶链反应(Polymerase cham reaction,PCR)扩增、变性聚丙烯酰胺凝胶电泳、硝酸银染色.分析D9S171微卫星多态性位点的LOH.结果头颈鳞癌原发癌组织的D9S171多态性位点的LOH发生率为62.96%;LOH发生与患者的临床分期有相关关系(P＜0.01).颈清扫淋巴结组织D9S171多态性位点的LOH发生率(35.60%),高于常规病检的阳性率(17.0%),(P＜0.01).结论D9S171多态性位点LOH与头颈鳞癌的发生有密切关系,LOH分析可能作为检测头颈鳞癌颈淋巴结转移的手段之一.

A study on the loss of heterozygosity at D9S171 locus in the primary tumor tissues and neck lymph nodes among cases with HNSCC

Objective To investigate the frequency of loss of heterozygosity(LOH)at D9S171 locus in the primary tumor tissues and lymph nodes taken from neck dissection among cases with head and neck squamous cell carinoma(HNSCC)and its clinical implications in these conditions. Methods The total DNA was extracted from primary tumor tissues and meck lymph nodes respectively by the standardized phenol-chloroform method. Then,polymerase chain reaction(PCR)was carried out to amplify D9S171 and the products were separated on 10% denatured polyacrylamide gels,with the gels stained by silver nitrate and LOH analyzed at this locus afterwards. Results The frequency of LOH at D9S171 was 62.96% in the primary tumor tissues,with a sigmificantly correlating tendency with the clinical staging for these cases(P<0.01). while this frequency was 35.60% in the lymph nodes collected from neck dissection, significantly higher than that found in routine histopathology(17.00%,P<0.01). Conclusions LOH at D9S171 closely correlated with the tumorogenesis of HNSCC and the analysis on this locus can be taken as one of the means to detect neck lymph metastasis from HNSCC.