Exploration Of The Molecular Mechanism Of HL-60 Cell Apoptosis Induced By Lycorine

Leukemia is one of the most threatening hematological malignant cancer today and chemotherapy remains the conventional method in treatment.Lycorine is one of alkaloids extracted from Amaryllidaceae family,showing a strong natural anti-cancer activity.This research focused on the expression change of several apoptosis-related proteins in HL-60 cell after treatment of lycorine in order to study the molecular mechanism of the anti-leukemia activity of lycorine.HL-60 cells were treated with 5.0μM lycorine for 24 h,with the group treated without lycorine as control.RT-PCR was used to analyze the influence of lycorine on the expression of TNF-αwith the total RNA extracted from the two groups.Western blotting was used to analyze the influence of lycorine on the expression of TNF-α,Bid,IκB,NF-κB and survivin with the proteins extracted from four groups of HL-60 cell, which were treated by 0μM,1.25μM,2.5μM and 5.0μM lycorine respectively for 24 hours.Finally,immunocytochemistry was used to analyze the nuclear import of NF-κB in two groups of HL-60 cell treated by 0μM and 5.0μM lycorine respectively for 24 hours.The results show that the expression of TNF-αwas up-regulated,Bid down-regulated, tBid up-regulated,IκB unchanged,p-IκB down-regulated,NF-κB nuclear import increased and survivin down-regulated with the increase of the concentration of lycorine. It is concluded that the activation of mitochondria apoptosis pathway indicated by the analysis of TNF-αand Bid cleavage,the inhibition of transcription regulation function of NF-κB in nucleus indicated by the analysis of IκB phosphorylaton and NF-κB nuclear import and the interference of IAPs indicated by the down-regulation of survivin were involved in the apoptosis of HL-60 cell induced by lycorine.