Asokan G S, Jeelani S, Gnanasundaram N
Associate Professor, Department of Oral Medicine and Radiology, Tagore Dental College and Hospital , Chennai, India .
Reader, Department of Oral Medicine and Radiology, Indira Gandhi Institute of Dental Science , Pondicherry, India .
J Clin Diagn Res. 2014 Oct;8(10):ZC09-12. doi: 10.7860/JCDR/2014/9251.4949. Epub 2014 Oct 20.
The present study was conducted to evaluate epigenetic alteration of five tumour suppressor genes in the oral precancer and cancer patients.
The study was carried out in three groups namely control group of five people (normal healthy individuals), 10 oral leukoplakia patients and 10 oral squamous cell carcinoma patients. Incisional biopsy was done and part of the tissue sent for histological examination and part of tissue sent for hypermethylation study of p16, p15, hMLH, MGMT, E-cadherin tumour suppressor genes. Methylation specific polymerase chain reaction was carried out for detecting methylation in promoter regions of tumour suppressor genes. The resultant PCR products were run in a 2.5% agarose gel and the promoter hypermethylation status of the five tumour suppressor genes were analysed.
In oral Leukoplakia patients, 60% of methylation in the case of p16 gene, 30% of methylation in the case of MGMT gene and 60% of methylation in the case of E-cadherin gene. In oral Squamous cell carcinoma patients, 60% of methylation in the case of p16 gene, 40% of methylation in the case of MGMT, 60% of methylation in the case of E-cadherin gene, 20% in case of p15,10% in case of hMLH gene.
Our results suggest that epigenetic mechanisms of inactivation of tumour suppressor genes, such as aberrant methylation of p16 and E-cadherin genes occur early in head and neck tumourigenesis and might play a role in the progression of these lesions.
本研究旨在评估口腔癌前病变和癌症患者中五个肿瘤抑制基因的表观遗传改变。
本研究分为三组,即五名正常健康个体组成的对照组、10名口腔白斑患者和10名口腔鳞状细胞癌患者。进行了切取活检,一部分组织送去做组织学检查,另一部分组织送去做p16、p15、hMLH、MGMT、E-钙黏蛋白肿瘤抑制基因的高甲基化研究。采用甲基化特异性聚合酶链反应检测肿瘤抑制基因启动子区域的甲基化情况。将所得的PCR产物在2.5%的琼脂糖凝胶中进行电泳,并分析五个肿瘤抑制基因的启动子高甲基化状态。
在口腔白斑患者中,p16基因甲基化率为60%,MGMT基因甲基化率为30%,E-钙黏蛋白基因甲基化率为60%。在口腔鳞状细胞癌患者中,p16基因甲基化率为60%,MGMT基因甲基化率为40%,E-钙黏蛋白基因甲基化率为60%,p15基因甲基化率为20%,hMLH基因甲基化率为10%。
我们的结果表明,肿瘤抑制基因失活的表观遗传机制,如p16和E-钙黏蛋白基因的异常甲基化,在头颈部肿瘤发生早期就会出现,并可能在这些病变的进展中发挥作用。