Effects of Kanglaite on apoptosis and apoptotic protein procaspase-3 and caspase-9 expression in hepatic carcinoma BEL-7404 cells

Objective:To examine the effects of Kanglaite injection(KLT) on proliferation,apoptosis,and apoptotic protein procaspase-3 and caspase-9 expressions in human hepatic carcinoma BEL-7404 cells.Methods:The inhibitory effect of KLT and positive control cisplatin(DDP) on the proliferation of BEL-7404 cells was evaluated using MTT assay.Hoechst 33258 fluorescent staining was used to observe the morphological changes of apoptotic cells.Flow cytometry was performed to determine the apoptotic rate.The expression levels of procaspase-3 and caspase-9 proteins were detected using Western blot analysis.Results:MTT assay showed that after BEL-7404 cells were treated with KLT 10,20,40,80,and 160 μL/mL,the apparent inhibition of cell proliferation was observed at 48 h after KLT 80 μL/mL.Hoechst 33258 staining revealed typical apoptotic morphological changes.FCM detection results indicated that the cell apoptotic rate was(1.23±0.40)% for control group,(32.53±0.65)% for DDP 10 μg/mL group,and(3.13±0.32)% for KLT 80 μL/mL(P0.01).Western blot analysis suggested that expression level of procaspase-3 was significantly decreased(P0.01),while the expression of caspase-9 was significantly increased(P0.01) at 48 h after treatment with KLT 80 μL/mL and DDP 10 μg/mL.Conclusion:KLT inhibits the proliferation and induced apoptosis of BEL-7404 cells by down-regulating procaspase-3 expression and up-regulating caspase-9 expression.