Effects of recombinant human tumor necrosis factor-alpha on HL-60 cells in vitro and in vivo.
- Author:
Xia LIU
1
;
Yuan-Zhong CHEN
;
Yong WU
;
Mei-Juan HUANG
;
Da-Liu YANG
;
Jiang-Rui GUO
Author Information
1. Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antineoplastic Agents;
pharmacology;
Apoptosis;
drug effects;
Cell Proliferation;
drug effects;
Dose-Response Relationship, Drug;
HL-60 Cells;
Humans;
Leukemia, Promyelocytic, Acute;
drug therapy;
Mice;
Mice, Inbred BALB C;
Mice, Nude;
Neoplasm Transplantation;
Recombinant Proteins;
pharmacology;
Tumor Necrosis Factor-alpha;
pharmacology
- From:
Journal of Experimental Hematology
2006;14(3):477-480
- CountryChina
- Language:Chinese
-
Abstract:
To study the effects of recombinant human tumor necrosis factor-alpha (rhTNF-alpha) on HL-60 cells in vitro and in vivo, MTT and colony forming assay were used to examine the effects of rhTNF-alpha on proliferation of HL-60 cells; AO/EB (acridine orange-ethidium bromide) staining, Annexin-V flow cytometry analysis and TUNEL assay were used to detect apoptotic cells. The effect of rhTNF-alpha on xenograft growth of HL-60 cells was evaluated by tumor inhibition rate, histology, ultrastructure and TUNEL assay. The results showed that rhTNF-alpha inhibited the proliferation of HL-60 cells in a dose-dependent manner. Staining of cells with AO/EB revealed that rhTNF-alpha induced nuclear chromatin condensation and fragmentation. Positive Annexin V-FITC on cell membrane showed that rhTNF-alpha induced apoptosis of HL-60 cells in a dose-dependent manner. TUNEL assay showed that the apoptotic percentage of HL-60 cells reached 37.5% when incubated with 3200 U/ml rhTNF-alpha for 48 hours. In vivo rhTNF-alpha inhibited xenograft growth of HL-60 cells with the highest inhibition rate of 60.33%. Pathologically it was found that there were necrotic areas in the tumors of groups treated with rhTNF-alpha. There were more apoptotic cells in treatment groups than in that control group by transmission electron microscopy (TEM) and TUNEL assay. It is concluded that rhTNF-alpha is able to inhibit the proliferation of HL-60 cells and to induce apoptosis of HL-60 cells in vitro and in vivo.
