目的:探讨年龄相关性白内障与正常透明晶状体前囊上皮细胞基因表达差异,为年龄相关性白内障发病机制的研究及治疗提供理论依据。方法:提取年龄相关性白内障病人晶状体和正常透明晶状体前囊中央上皮细胞RNA,分别用cy3、cy5标记,再与含8064个基因点的芯片杂交,筛选两个样本间表达差异的基因,并对这些基因进行初步功能分析。按不同功能类型分别选择9个存在显著表达差异的基因,应用实时荧光定量PCR技术验证基因芯片可靠性。结果:年龄相关性白内障前囊中央上皮细胞中有724个差异表达基因,包括438个表达下调基因和286个表达上调基因。进行初步功能分类分析发现这些表达差异基因主要涉及晶状体结构蛋白基因、细胞骨架蛋白基因、细胞增殖及细胞凋亡以及细胞应激反应等方面。结论;年龄相关性白内障与透明晶状体前囊上皮细胞存在基因表达差异,其中多数基因呈表达下调。表达下调基因功能涉及的生物学过程导致晶状体维持内环境稳定的能力下降,形成白内障。
AIM: To identify the differences of gene expression between human age - related cataract ahd clear lenses. METHODS: The RNA were extracted from human age related cataract and clear lens epithelial cells, labeled with cy3/cy5 as probes, then were hybridized to cDNA chip containing 8 064 genes. The differential expressions of the genes were screened. Furthermore, a primary classification of these genes function was given. The expression levels of the identified genes were further evaluated by real time polymerase chain reaction. RESULTS: 286 genes expression were observed to increase and 438 genes expression were observed to decrease in cataractous lens epithelial cells as compared with normal lens. According to functional analysis, the changed genes in cataract lens are associated with lens structural components, cytoskeleton, cell cycle, apoptosis and stress responses. CONCLUSION: These data suggest that there are differences in gene expression between cataract and clear human lens epithelial cells. The majority of genes changed in cataract exhibited decreased expression. Processes associated with the down - regulated genes may reflect the inability of the lens to maintain its homeostasis and transparency.