MS-HRM技术定量检测喉鳞癌Syk(L)基因启动子甲基化的方法及意义

Quantitative determination for methylation level Syk( L) promoter by MS-HRM in human laryngeal squamous cell carcinoma and its clinical significance

目的利用MS-HRM技术建立Syk（L）基因启动子甲基化定量检测的方法,并探讨其甲基化程度与喉鳞癌临床病理参数的相关性。方法提取48份喉鳞癌组织及20份癌旁正常喉黏膜组织DNA并进行甲基化修饰。用MSHRM技术定量检测Syk（L）基因启动子甲基化水平,并分析评价其与喉鳞癌临床病理参数的相关性。结果 MS-HRM检测Syk（L）基因启动子甲基化结果为20份癌旁正常喉黏膜组织均位于0%区域;喉鳞癌组织中6份位于0%区域,8份位于0%-30%区域,20份位于30%-70%区域,14份位于70%-100%区域。Syk（L）基因启动子甲基化水平与喉鳞癌临床分型差异无统计学意义（P〉0.05）,但与临床T分期、病理组织学分级、颈淋巴结转移差异有统计学意义（P〈0.05）,且Syk（L）甲基化水平随临床T分期及病理组织学分级增高而增高。结论 MS-HRM定量检测Syk（L）基因启动子甲基化有望成为喉鳞癌进展评估分子指标检测的有效手段。

Objective To establish quantitative method for detection of methylation level of syk（ L） promoter with the methylation seusitive-high resolution melt（ MS-HRM） technology and to its association with pathological parameters in human laryngeal squamous cell carcinoma. Methods 48 human laryngeal squamous cell carcinoma tissues samples and 20 adjacent normal tissues samples were enrolled into this study. DNA was extracted and modified for methylation deteemination. The Syk（ L）methylation levels in laryngeal squamous cell carcinoma tissue samples were detected with MS-HRM. The relationship between the methylation statues of Syk（ L） and clinical classification,clinical T stage,histopathological grading,1ymph node metastasis was analyzed. Results The methylation levels of 20 adjacent normal tissues samples and 6 human laryngeal squamous cell carcinoma tissues samples were overlapped with 0% methylated calibrator at the Syk（ L） promoter region by MS-HRM. The methylation levels of 8 cancer samples ranged between 0% and 30%. The methyltion levels of 20 cancer samples ranged between30% and 70%. The methylation levels of 14 cancer samples ranged between 70% and 100%. There was no significant association between methylation levels of Syk（ L） and clinical classification（ P 〉0. 05）. The methylation levels of Syk（ L） was correlated with the clinical T stage,histopatho-logy grade and lymphatic metastasis（ P〈 0. 05）. Conclusion Syk（ L） methylation MS-HRM quantitative methods have many promising applications in the molecular estimation of malignancy in human laryngeal squamous cell carcinoma.