[Effect of bone marrow stromal cells on the apoptotic sensitivity of HL-60 and HL-60/VCR cells].
Rong Liang, Gao-Sheng Huang, Zhe Wang, Xiequn Chen, Qing-Xian Bai, Weiping Zhang, Juanhong Wang, Wenqing Wang, Ying Guo
Abstract
Rong Liang, Gao-Sheng Huang, Zhe Wang, Xiequn Chen, Qing-Xian Bai, Weiping Zhang, Juanhong Wang, Wenqing Wang, Ying Guo
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.
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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.
Key concepts: Molecular biology, Apoptosis, Stromal cell, Bone marrow, Cell culture, Acridine orange, Chemistry, Staining