Chinese Journal of Cancer Prevention and Treatment
OBJECTIVE: To investigate the effect on Caspases expression in PGCl_3 cells for studying mechanism of apoptosis of PGCl_3 cells induced by death associated protein kinase(DAPK). METHODS: Eukaryotic express vector pcDNA3.1-DAPK was transfected into high-metastasis non-small lung cancer cell PGCl_3. Morphologic assessment of apoptosis was performed with fluorescence microscope. Subdiploid peak of DNA, cell cycle and mitochondrial membrane potential were analyzed by flow cytometry. Changes of Caspase-3, Caspase-6 and Caspase-8 expressions were detected by RT-PCR. RESULTS:The pcDNA3.1-DAPK-transfected could induce apoptosis of PGCl_3 cells, but it didn not affect on PGCl_3 cells cycle and mitochondrial membrane potential, t= 1.816,P=0.328. Caspase-3 and Caspase-8 expressions were upregulated, and meanwhile Caspase-6 did not change in apoptosis of PGCl_3 cells induced by DAPK. CrmA (100 μmol/L), a Caspases inhibitor, didn’t suppress cell apoptosis, t=0.969, P= 0.423. CONCLUSIONS: Apoptosis of PGCl_3 cells induced by over-expression of DAPK may be associated with Caspase-8 and Caspase-3 pathway. Caspase-6 may not be an important pathway. Additionally, apoptosis of PGCl_3 cells induced by DAPK may have another independent pathway at the same time.