Zhang Gang, Fu Jile, Luo Shaojun, Tang Shaoming, Liang Lie
Institute of Plastic Surgery, Guangdong Medical College, Zhanjiang Guangdong 524001, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008 Nov;22(11):1311-3.
To study the mutations at 1 573 fragment of TNF receptor II (TNFR-II) gene in patients with keloid.
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.
All the concentration of the extracted DNA in trial were higher than 0.50 microg/microL 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.
There is a correlation between the mutation at 1 573 fragment of TNFR-II gene and keloid.
研究瘢痕疙瘩患者肿瘤坏死因子受体II（TNFR-II）基因1573片段的突变情况。
从22例患者（男6例，女16例，年龄18 - 53岁）捐赠的22份瘢痕疙瘩组织样本中提取组织DNA，所有瘢痕疙瘩均经病理学家检查并分类。从同一患者提取外周血DNA作为对照。采用聚合酶链反应（PCR）从瘢痕疙瘩组织DNA和外周血DNA中扩增TNFR-II基因的1573片段。对PCR产物直接测序，然后与基因库数据进行比较。
试验中提取的所有DNA浓度均高于0.50μg/μL，提取DNA的纯度（A260/A280）均高于1.5。经琼脂糖凝胶电泳检测，其大小接近设计的DNA片段，符合试验要求。22例瘢痕疙瘩中有13例检测到TNFR-II基因1573片段突变，突变发生率为59.1%。其中，9例在密码子1663处发生点突变，占40.9%。所有外周血样本均未检测到TNFR-II基因突变。瘢痕疙瘩DNA与外周血DNA之间存在显著差异（P < 0.01）。突变涉及点突变、缺失、插入以及多位点和多类型。
TNFR-II基因1573片段突变与瘢痕疙瘩之间存在相关性。
