PURPOSE To investigate apoptosis induced by hydroxycamptothecin (HCPT) in human Tenon’s capsule using fibroblasts cultured from human Tenon’s capsule (HTFs), and to investigate the mechanism of induction. METHODS HTFs were treated with 0-4 mg/L HCPT for 24 h. Cell proliferation was measured by MTT, and apoptotic cells were analyzed by Hoechst 33258 stain. The mRNA and protein levels of caspase-3, -8, and -9 were detected by RT-PCR and western blotting. RESULTS By MTT assay, HCPT induced apoptosis in HTFs in a concentration- and time-dependent manner. Hoechst 33258 staining and transmission electron microscopy showed typical apoptotic morphology such as condensed chromatin, irregular nuclei, and apoptotic body formation. The mRNA and protein levels of caspase -3 and caspase-9 were upregulated, while caspase-8 was unchanged. Z-VAD-FMK inhibited the apoptosis of fibroblasts induced by HCPT. The expression levels of caspase-3 and caspase-9 were downregulated after Z-VAD-FMK treatment. CONCLUSION Caspase-3 and caspase-9 are important elements in regulating HCPT-induced apoptosis in HTFs. |