Degeneration and regeneration of normal conjunctival epithelium. Role of apoptosis-associated genes

Background. The aim of the present study was to evaluate the age-dependent physiological pattern of apoptosis-related genes in the normal conjunctiva. Patients and methods. Healthy probands (group A: 24 probands and group B: 24 probands) were subjected to modified impression cytology using a PTFE membrane for evaluation of age-dependent expression of apoptosis-relatecl genes in the normal conjunctiva RNA was isolated from the cytologic impression specimens and used for cDNA synthesis employing Super-ScriptTM II reverse transcriptase. Results. RT-PCR revealed the detection of apoptosis-related mRNAs as follows (group A compared to group B): Apaf-1: 4.2/4.2%, Bcl-2:41.7/41.7%, Bim: 0/0%, Bag-1: 70.8/100%, p53: 0/8.3%, Casp-3:4.2/66.7%, Casp-5:70.8/37.5%, Casp-8:62.5/20.8%, Casp-9:25/12.5%, c-myc: 83.3/95.8%, BAX: 91.7/70.8%, and BAD: 95.8/87.5%. Conclusion. The results revealed an age-dependent expression of apoptosis-associated genes (proapoptotic: Casp-3, Casp-5, Casp-8, antiapoptotic Bag-1) in normal human conjunctiva. Taken together these investigations provide basic information to understand the expression of apoptosis associated genes in various diseases of the conjunctiva.