瘢痕疙瘩TNF受体Ⅱ基因1573位点突变的研究

RELATIONSHIP BETWEEN MUTATION AT 1 573 FRAGMENT OF TNF RECEPTOR II GENE AND KELOID

目的探讨瘢痕疙瘩患者TNF受体Ⅱ(TNF receptorⅡ,TNFR-Ⅱ)基因1573位点突变的情况。方法收集22例自愿捐献的经临床及病理确诊的瘢痕疙瘩标本,其中男6例,女16例;年龄18～53岁。设患者自身外周静脉血标本为正常对照。提取基因组DNA,PCR扩增TNFR-Ⅱ基因1573位点片段,DNA测序,将测序结果与GeneBank比较。结果实验提取DNA浓度均>0.5μg/μL,纯度(A260/A280)均>1.5,经琼脂糖凝胶电泳检测,与所设计DNA片段大小相近,符合实验要求。13例瘢痕疙瘩标本检测示不同程度突变,突变率为59.1%;9例1663编码子发生点突变,占总数的40.9%。与外周静脉血比较,差异均有统计学意义(P<0.01)。突变类型主要为点突变、插入、缺失,为多位点、多类型,呈多态性。结论TNFR-Ⅱ基因1573位点突变与瘢痕疙瘩的发生有关。

Objective To study the mutations at 1 573 fragment of TNF receptor Ⅱ （TNFK-Ⅱ） genem patients with keloid. Methods The tissue DNA was extracted from 22 samples of keloids donated by 22 patients （6 males and 16 females, aged 18-53 years）, and all keloids were examined and classified by pathologist. The peripheral blood DNA was extracted from the same patients as the control. PCR was used to amplify the 1 573 fragment of TNFR-Ⅱ gene from the keloid tissue DNA and peripheral blood DNA. The PCR products were sequenced directly and then compared with the GeneBank data. Results All the concentration of the extracted DNA in trial were higher than 0.50 μg/μL and the purity （A260/A280） of the extracted DNA were higher than 1.5. It closed to the magnitude of the design DNA fragment by agarose gel electrophoresis examining, and corresponded with the test requirement. Mutations at 1 573 fragment of TNFR-Ⅱ gene were detected in 13 out of 22 keloids. The mutation incidence was 59.1%. Among them, 9 had point mutation at codon 1 663, accounting 40.9%. No TNFR-Ⅱ gene mutation was detected in all peripheral blood samples. There were significant difference between keloids DNA and peripheral blood DNA （P 〈0.01）. The mutations involved point mutation, deletion and insertion as well as multisite and multitype. Conclusion There is a correlation between the mutation at 1 573 fragment of TNFR-Ⅱ gene and keloid.