人角膜基质细胞中cyclin—CDK—CKI网络成员的表达

Expressions of cycling-CDK-CKI network members in human corneal keratocytes.

为进一步调控角膜基质细胞周期、抑制角膜基质细胞增殖提供基础和靶点 ,检测原位人角膜基质细胞和体外培养的人角膜基质细胞是否表达细胞周期蛋白 (cyclin)—细胞周期蛋白依赖性激酶 (cyclin dependentkinase,CDK)—细胞周期蛋白依赖性激酶抑制因子 (cyclin dependent kinase inhibitor,CKI)网络成员。方法 :制作人角膜冰冻切片 ,并体外培养人角膜基质细胞 ,分别用免疫组织化学和免疫细胞化学的方法检测其中 cyclin D、E,cdk2、4,p2 1WAF1的表达。结果 :体外培养的人角膜基质细胞中有 cyclin D、E,cdk2、 4,p2 1WAF1蛋白水平的阳性表达 ,而原位人角膜基质细胞中无阳性表达。结论 :体外培养的人角膜基质细胞有别于静止状态的角膜基质细胞 ,与在体激活的角膜基质细胞相似 ,cyclin—CDK—CKI成员在其中有阳性表达的结果 ,为进一步通过人为下调 cyclin/ CDK水平、上调

Objective: To provide basis and targets for regulation of cell cycle and inhibition of cell proliferation. To definite the expressions of cyclin-cyclin dependent kinase (CDK)-cyclin dependent kinase inhibitor (CKI) network members in cultured human corneal keratocytes and in situ human corneal keratocytes. Methods: Human corneal cryosection was made, and human corneal keratocytes was cultured in vitro, then with immunohistochemistry the expressions of cyclinD, E. CDK2. 4, p21WAF1 were detected. Results: Cultured human corneal keratocytes express cyclinD, E, CDK2, 4, p21WAF1 in protein level, but there was no positive expression in corneal keratocytes in situ. Cultured human corneal keratocytes is like activated keratocytes in vivo. Conclusions: Based on the different expressions between cultured keratocytes and corneal keratocytes in situ, the results provide basis for our further study to inhibit proliferation of keratocytes through downregulating cyclin/CDK level and upregulating CKI level.