Effect of bone marrow stromal cells on the apoptotic sensitivity of HL-60 and HL-60/VCR cells.
- Author:
Rong LIANG
1
;
Gao-Sheng HUANG
;
Zhe WANG
;
Xie-Qun CHEN
;
Qing-Xian BAI
;
Wei-Ping ZHANG
;
Juan-Hong WANG
;
Wen-Qing WANG
;
Ying GUO
Author Information
1. Department of Hematology, The Fourth Millitary Medical University, Xi'an 710032, China. bettyliang9845@hotmail.com
- Publication Type:Journal Article
- MeSH:
Antineoplastic Agents;
pharmacology;
Apoptosis;
drug effects;
physiology;
Blotting, Western;
Bone Marrow Cells;
cytology;
physiology;
Caspase 3;
metabolism;
Cell Cycle;
drug effects;
physiology;
Cell Line;
Coculture Techniques;
Dose-Response Relationship, Drug;
Drug Resistance, Neoplasm;
Fibroblasts;
cytology;
physiology;
Flow Cytometry;
HL-60 Cells;
Humans;
Proto-Oncogene Proteins c-bcl-2;
metabolism;
Stromal Cells;
cytology;
physiology;
Topotecan;
pharmacology;
Vincristine;
pharmacology
- From:
Journal of Experimental Hematology
2005;13(2):286-292
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the effects of human bone marrow fibroblastoid stromal cell line (HFCL) on chemosensitivity of acute myeloid leukemia sensitive HL-60 cell line and multidrug-resistant (MDR) HL-60/VCR cell line in vitro co-culture. Setting up co-culture system of HL-60 or HL-60/VCR cells in direct contact with HFCL cells, or with HFCL cells separated by transwell, and exposing HL-60 or HL-60/VCR cells to different concentrations of topotecon (TPT), morphologic evidence for apoptosis was determined by staining with Wright-Giemsa stain and acridine orange/ethidium bromide (AO/EB). Cell cycle, sub-G(1) and annexin V FITC staining were detected by flow cytometry. The expression of active caspase-3, Bcl-2 and Pgp was detected by Western blot. The results showed that HL-60 or HL-60/VCR cells treated by TPT revealed characteristic apoptotic morphological changes by Wright-Giemsa and AO/EB staining. The percentage of annexin V-positive cells and apoptotic cells decreased when they were cocultured with HFCL cells. The proportion of G(0)/G(1) HL-60 or HL-60/VCR cells treated by TPT increased and the sub-G(1) appeared significantly, but apoptotic and sub-G cells reduced after direct contact with HFCL cells. Meanwhile, although HL-60 or HL-60/VCR cells treated by TPT expressed activated caspase-3, and the expression of Bcl-2 decreased, the expression of activated caspase-3 decreased and Bcl-2 increased after direct contact with HFCL cells. In conclusion, HFCL stromal cells can prevent TPT-induced apoptosis in HL-60 and HL-60/VCR cells via modulation of Bcl-2 and active caspase-3.