Apoptosis of HL-60 Cells Induced by Peptide from Hemocyte of Horseshoe Crab
10.3321/j.issn:1000-467X.2001.06.012
- VernacularTitle:鲎血细胞多肽诱导HL-60细胞凋亡
- Author:
Hai-Tao ZHANG
1
;
Kang-Rong CAI
;
Qi-Feng ZHU
Author Information
1. Guangdong Medical College
- From:Chinese Journal of Cancer
2001;20(6):616-619
- CountryChina
- Language:Chinese
-
Abstract:
Objective：This study was designed to investigate the apoptosis of HL 60 cells induced by the peptide from hemocyte of horseshoe crab (horseshoe crab hemocyte peptide). Methods： Cytotoxicity and cell viability were assayed by MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope; the apoptosis of the cells was confirmed by flow cytometry; cell cycle was analyzed by flow cytometry, changes of cell membrane were observed by scanning electron microscope. Results： HL 60 cells treated with horseshoe crab hemocyte peptide were showed apparently cytotoxicity with IC50 24 μ g/ml. The morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed with fluorescence microscope. Treated with horseshoe crab hemocyte peptide 50- 100 μ g/ml for 6 h, apoptostic HL 60 cells were predominant. However the HL 60 cells were shown necrotic and apoptostic cells were reduced when treatment with horseshoe crab hemocyte peptide of 200 μ g/ml. SubG1 peak was detected by flow cytometry. Cell cycle analysis showed Phase G1 cells decreased and Phase G2 cells increased with horseshoe crab hemocyte peptide 50 μ g/ml treating for 0- 12 h. Ratio of apoptosis increase depended on concentrations of horseshoe crab hemocyte peptide in 25- 100 μ g/ml but it reduced in 200 μ g/ml, the HL 60 cells were shown necrotic mainly. Scanning electron microscope showed membrane of HL 60 cells was damaged by horseshoe crab hemocyte peptide treatment,cell membrane showed some holes. Conclusion： Peptide from hemocytes of horseshoe crab can induce apoptosis in HL 60 cell,it was early event,and may correlate with membrane damage. The HL 60 cell in Phase G1 seemed to be sensitive to the peptide.
