Apoptosis And Mechanism Of Human Tenon’s Capsule Fibroblast Induced By Hydroxycamptothecin

Purpose. To investigate apoptosis induced by hydroxycamptothecin (HCPT) inhuman Tenon’s capsule using fibroblasts cultured from human Tenon’s capsule(HTFs), and to investigate the mechanism of induction.Methods. The fibroblasts cultured from human Tenon’s capsule (HTFs) were treatedwith0-4mg/L HCPT for24h. The cell proliferation was measured by MTT, andapoptotic cells were analyzed by Hoechst33258stain. The mRNA and proteinexpression levels of caspase-3,-8,-9were detected by RT-PCR and Western blotting,respectively.Results. According to the MTT assay, HCPT induced human Tenon’s capsulefibroblasts apoptosis in a concentration-and time-dependent manner. Hoechst33258staining and transmission electron microscopy showed typical apoptotic morphologysuch as condensed chromatin, irregular nuclei, and apoptotic body formation. ThemRNA and protein expression levels of caspase-3, and-9were up-regulated, whilecaspase-8was no change. Z-VAD-FMK, caspase inhibitor, inhibited the apoptosis offibroblasts induced by HCPT. The expression levels of caspase-3, and-9weredown-regulated after Z-VAD-FMK treatment.Conclusions. Caspase-3and-9are important elements regulating the HCPT-inducedapoptosis in human Tenon’s capsule fibroblasts.