新基因c24的克隆及其在正常及白内障晶状体上皮细胞中的表达  

Cloning of a novel gene c24 and its expression in lens epithelium cells of normal and age-related cataractous eye

作　　者：刘燕[1] 周健[2] 刘新平[3] 药立波[3] 惠延年[2] 

机构地区：[1]第四军医大学唐都医院眼科,西安710038 [2]第四军医大学西京医院眼科,西安710032 [3]第四军医大学基础部生物化学与分子生物学教研室,西安710033

出　　处：《眼科研究》2007年第11期801-804,共4页Chinese Ophthalmic Research

基　　金：国家自然科学基金(30000185);第四军医大学唐都医院苗子人才基金资助

摘　　要：目的克隆新基因c24并研究其在正常晶状体和年龄相关性白内障晶状体上皮细胞(LECs)中的表达,并比较二者表达的差异。方法用改良的消减杂交法克隆新基因c24,地高辛标记c24cDNA片断制作探针,用原位杂交法研究其在晶状体前囊膜切片上的表达,并进行统计学处理。结果克隆到了新基因c24的cDNA片断,长约532bp,c24在正常晶状体和年龄相关性白内障LECs中均有表达;其在年龄相关性白内障LECs中的表达高于其在正常LECs的表达。结论c24在正常晶状体和年龄相关性白内障LECs中均有表达;在年龄相关性白内障LECs中,其表达较正常晶状体增高,提示其可能在白内障的发生中起一定的作用。Objective Previous studies suggested that cataract is associated with the altered expression of specific genes in the lens epithelium cells. In our previous study,we found c24,a novel gene from the subtractive cDNA library of the age-related cataract and normal lens, In this study ,we tried to clone the gene c24 and study its expression in lens epithelium cells（LECs） of normal and age-related cataractous eyes. Methods c24 was cloned by improving subtractive hybridisation,and the fragment of c24 was labelled by digoxin and hybridised by in situ hybridisation on the sections of capsule from normal group（ 10 cases from eyeball after cornea grafting） and cataractous group （10 cases from cataract surgery）. Results c24 was cloned successfully with the length 532 bp. c24 was expressed in LECs both age-related cataractous lens and normal lens, showing brown-blue particles in cytoplasm for NBT and BCIP and the positive cell rate was（80.3 ± 14. 3）% and （62.7 ± 15.6）% respectively with a significant difference （P 〈 0.05 ）. Conclusion c24 may play an important role in the etiology of age-related cataract.

关 键 词：新基因c24 年龄相关性白内障 晶状体上皮细胞 

分 类 号：R776.1[医药卫生—眼科] Q7[医药卫生—临床医学]