重庆地区开角型青光眼患者及其亲属和正常人群中小梁网糖皮质激素诱导反应蛋白基因突变的研究

Study on the gene mutation of TIGR in primary open angle glaucoma,their relatives and normal controls in Chongqing

目的 研究开角型青光眼患者及其亲属和正常人群中小梁网糖皮质激素诱导反应蛋白(TIGR)基因突变的情况。方法 (1)应用聚合酶链反应 (PCR)方法 ,对 15例开角型青光眼患者及其10例一级亲属和 2 0例正常对照者的TIGR基因的 3个外显子、部分内含子及部分启动子 (7对引物 )的各个片段进行扩增 ,应用单链空间构象多态 (SSCP)分析法 ,筛选可能存在突变的PCR产物。 (2 )将筛选出的PCR产物交上海基康公司测序。 (3)对突变片段行生物信息学分析。结果 (1)在重庆地区 15例原发性开角型青光眼患者中 ,共发现TIGR基因突变者 5例 ,其中青少年型青光眼 4例 ;在 10例青光眼患者亲属中发现TIGR基因突变者 2例 ;正常对照者 2 0例中未发现TIGR基因突变。 (2 )PCR产物测序共发现 4个序列改变 ,其中编码区 2个 ,非编码区 2个。编码区的 2个突变位点(Ser5 5Thr、Asp2 4 7Stop)及第二内含子区bp35c→t的突变 ,均未见文献报道 ;而在启动子区域bp - 83c→t的突变有文献报道为 1个多态位点。 (3)生物信息学分析结果显示编码区的突变可导致氨基酸序列、蛋白质的二级结构及等电点、抗原结合位点等发生改变。结论 TIGR基因突变与青少年开角型青光眼的发生密切相关 ,由此可推测青光眼患者的亲属发病率较正常人高。TIGR基因突变可?

Objective To study trabecular meshwork induced glucocorticoid response protein (TIGR) gene mutation in primary open angle glaucoma (POAG), their relatives and normal controls in Chongqing. Methods (1) The coding sequence of TIGR was screened for sequence alterations using polymerase chain reaction (PCR) followed by single-strand conformation polymorphism (SSCP). (2) Samples corresponding to bands of altered mobility were sequenced. (3) The sequence alterations were analyzed by bioinformatics. Results (1) In 15 POAG, TIGR gene mutation was found in 5 cases. In 10 POAG relatives, 2 TIGR gene mutations were found. No TIGR gene mutation was found in 20 normal persons in Chongqing. (2) The mutation sites Ser55Thr, Asp247Stop and in 49021 c-t alteration discovered in the present study have not been reported previously. The 63155 c-t alternation has been reported as a polymorphism site previously. (3) Bioinformatics analysis indicated that the mutations in structural domain could lead to the changes in the amino acid sequence coded, second structure of protein, isoelectric point and antigen binding site. Conclusions We found that the TIGR gene mutations is closely related to the occurrence of JOAG in Chongqing. The relatives of POAG patients may have a higher incidence of TIGR gene mutation as compared with the controls. Furthermore, after analyzed by bioinformatics, we found that the alteration of structure and biological activity of TIGR protein resulted from gene mutation may be one of the important changes that leading to the occurrence of POAG.