DNA修复基因在核型年龄相关性白内障患者晶状体上皮细胞中的表达

Expression of DNA repair genes in lens epithelial cells of patients with age-related nuclear cataract

目的探讨DNA修复基因在核型年龄相关性白内障(ARC)患者晶状体上皮细胞(LECs)中的表达。方法从30例核型ARC患者(观察组)与30例玻璃体视网膜病患者(对照组)中分别随机选取3例患者,采用人DNA修复基因TaqMan表达芯片行预实验检测DNA修复基因的表达。对表达差异在1.5倍以上的基因在两组剩余患者中采用实时荧光定量聚合酶链式反应(RT-PCR)验证,并采用Western blot法检测其相应蛋白表达。结果与对照组相比,观察组LECs中有7个DNA修复基因(FANCD2、LIG1、MGMT、MSH2、RPA2、SMUG1、XRCC6)mRNA表达下调1.5倍以上(P<0.05),1个基因(TREX1)mRNA表达上调1.5倍以上(P<0.05)。RT-PCR验证结果显示,与对照组相比,观察组LECs中FANCD2、LIG1、MGMT、MSH2、RPA2、SMUG1、XRCC6mRNA表达水平分别下降1.89、2.11、2.25、1.97、1.86、2.05、2.33倍(P<0.05),TREX1mRNA表达上升2.18倍(P<0.05);Western blot检测相应蛋白表达与其变化一致。结论在核型ARC患者和对照人群LECs中,部分DNA修复基因的表达存在差异,这些基因功能失调可能在核型ARC的形成和发展中起到一定作用。

Objective To explore the expression of DNA repair genes in lens epithelial cells（LECs）of patients with age-related nuclear cataract.Methods Thirty age-related nuclear cataract patients（group A）and thirty controls（group B）were included in the study.As the preliminary experiments,human DNA repair mechanism TaqMan arrays plates were used to examine the mRNA expression of DNA repair genes in LECs of three age-related nuclear cataract patients in group A and three controls in group B.In the remaining patients of two groups,RT-PCR and Western blot were used to confirm the mRNA and protein expressions of the genes identified by the arrays plates and showed a expression difference of more than 1.5time.Results Compared with group B,the mRNA expressions of seven DNA repair genes（FANCD2,LIG1,MGMT,MSH2,RPA2,SMUG1 and XRCC6）in group A were down-regulated more than 1.5times and one gene（TREX1）mRNA expression was up-regulated more than 1.5times（P〈0.05）.RT-PCR showed that,compared to group B,the mRNA expressions of FANCD2,LIG1,MGMT,MSH2,RPA2,SMUG1 and XRCC6in LECs of group A were decreased by 1.89,2.11,2.25,1.97,1.86,2.05 and 2.33 times,respectively,and TREX1 mRNA expression was increased by 2.18 times（P〈0.05）.The changes in the expressions of the genes mentioned above were accordant with those reported by Western blot.Conclusion The expression differences of some DNA repair genes in LECs exist between age-related nuclear cataract patients and healthy people,which may have a certain effect on the occurrence and development of age-related nuclear cataract.