[Effect of bone marrow stromal cells on the apoptotic sensitivity of HL-60 and HL-60/VCR cells].
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.
Liang
Rong
R
Department of Hematology, The Fourth Millitary Medical University, Xi'an 710032, China. bettyliang9845@hotmail.com
Huang
Gao-Sheng
GS
Wang
Zhe
Z
Chen
Xie-Qun
XQ
Bai
Qing-Xian
QX
Zhang
Wei-Ping
WP
Wang
Juan-Hong
JH
Wang
Wen-Qing
WQ
Guo
Ying
Y
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D019772:Topotecan / Q000494:pharmacology; D014750:Vincristine / Q000494:pharmacology*
D000970:Antineoplastic Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D014750:Vincristine; D019772:Topotecan; D053148:Caspase 3