Ⅰ型胶原基因的克隆及其在正常及年龄相关性白内障晶状体上皮细胞的表达

Cloning of collagen typeⅠgene and its expression in lens epithelium in normal and age-related cataractous people

目的:克隆Ⅰ型胶原基因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Ⅰ型胶原基因,地 高辛标记Ⅰ型胶原cDNA片段制作探针,用原位杂交法研究 其在晶状体前囊膜切片上的表达,并作统计学处理.结果:克 隆到了Ⅰ型胶原基因的cDNA片段,长约600bp,Ⅰ型胶原在 正常晶状体和年龄相关性白内障晶状体上皮细胞中均有表 达;阳性细胞百分率分别为(73.0±14.9)%和(75.0± 16.3)%,阳性表达的积分指数分别为(0.60±0.15)和 (0.73±0.16),经统计分析,P>0.05,其在年龄相关性白内 障晶体上皮细胞的表达与在正常晶体上皮细胞的表达未见明 显差异.结论:Ⅰ型胶原在正常晶状体和年龄相关性白内障 晶状体上皮细胞中均有表达;在年龄相关性白内障晶状体上 皮细胞中,其Ⅰ型胶原的表达较正常晶状体未见减少,提示?

AIM : To clone the gene of collagen type Ⅰ, to study its expression in lens epithelium of normal and age-related cataractous people, and to compare the difference of their expression. METHODS: The collagen type Ⅰ gene was cloned by the improved subtractive hybridization, and the fragment of collagen type Ⅰ cDNA was labeled by digoxin and was hybrided by in situ hybridization on the section of capsule of normal and cataractous lens. RESULTS : Collagen type I gene expressed both in the normal and the age-related cataractous lens epithelium. The positive cell rate was respectively (73±14.9)% and (75±16.3)%, the positive signals index was respectively(0.60±0.15) and (0.73±0.16) and the expression of the two groups had no significant statistics difference. CONCLUSION : Collagen type Ⅰ gene is expressed in the normal and the age-related cataractous lens epithelium. There is no significant differences in the expression of collagen type Ⅰ gene in the two groups, indicating that collagen type Ⅰ gene may play an important role in the etiology of cataract and the formation of posterior capsule opacification after cataract surgery.