NOVEL TETRACYCLINE TIGECYCLINE EFFECTS ON HEMATOLOGICAL MALIGNANCIES VIA CASPASE 3,9 PATHWAY AND NFKB PATHWAY.
(Abstract release date: 05/21/15)
EHA Library. Shigemi H. 06/12/15; 102855; PB1633
Disclosure(s): University of Fukui, Faculty of Medical ScienceHematology and Oncology
Hiroko Shigemi
Contributions
Contributions
Abstract
Abstract: PB1633
Type: Publication Only
Background
Novel tetracycline derivative tigecycline has an antibiotic effect against a wide range of microorganisms including an activity in multi-resistant drug infections and modulates the production of cytokines and chemokines. FDA approved it for the treatment of complicated infections in 2005. Besides it might affect hematological malignancies. Skrtic et al. reported tigecycline’s anti-leukemia activity in TEX cell lines and M9-ENL-1 cell lines. The phase I clinical trial has undergone in USA.
Aims
We examined the anti-leukemic effect and elucidated the mechanism of tigecycline. Furthermore, we confirmed tigecycline is also effective on cytarabine resistant (AraC-R) HL60 cell line and clofarabine resistant (CAFdA-R) HL60 cell line. Two cell lines showed 200-fold and 100-fold resistant to parental HL60 cells, respectively.
Methods
To assess the effects of tigecycline on cell viability in acute myeloid leukemia HL60 cell line, acute lymphocytic leukemia CEM cell line, multiple myeloma U266 cell line and chronic myeloid leukemia K562 cell line, XTT assay was performed. Annexin-PI assay, sub G1 analysis in cell cycle were performed using by flowcytometry in HL60 and CEM cell line. Apoptosis related protein (caspase 3, 8, 9, Bax, Bad, Bclxl, Bcl2) and the phosphorylation of IKKα/β, NFκB were detected by Western Blotting. The effectiveness of tigecycline on AraC-R HL60 cell line and CAFdA-R HL60 cell line, those have been established in our laboratory, were evaluated. Sensitivities on Bcl2 inhibitor (ABT737) and NFkB inhibitor (BAY11-7821) in HL60, CEM, AraC-R HL60 and CAFdA-R HL60 cells, were also examined.
Results
1) The cell viability was measured at 72 hours by XTT assay. The values of Inhibition Concentration 50% (IC50) of tigecycline in HL60, CEM, K562 and U266 cells were 5.0μM, 2.5μM, 2.7μM and 21.2μM respectively. It was shown that tigecycline was cytotoxic to leukemic cells. 2) Tigecycline induced apoptosis in a dose- and time- dependent manner. In addition, sub G1 analysis showed that tigecycline suppressed S phase in cell cycle. 3) Tigecycline suppressed the expression Bcl2 in a time-dependent manner and inhibited the phosphorylation of NFκB in HL60 cells and CEM cells. Tigecycline activated caspase3, 9. 4) We examined the effectiveness of tigecycline on AraC-R HL60 cells and CAFdA-R HL60 cells. The values of IC50 of tigecycline were 6.7μM (AraC-R) and 2.0μM (CAFdA-R). ABT737 and BAY11-7821 displayed more effective on resistant cells compared to HL60 parental cells. The values of IC50 of ABT737 were 881nM (Parental), 73.3nM (AraC-R), 68.4nM (CAFdA-R). Those of BAY11-7821 were 2.4μM (Parental), 1.1μM (AraC-R), 1.5μM (CAFdA-R).
Summary
Tigecycline showed an anti-leukemic effect on HL60, CEM and K562 cell lines. Tigecycline is presently used for the treatment of certain infections. Pharmacological concentration of tigecycline is useful for treatment of cancer and leukemia. Targeting caspase3, 9 pathway and NFκB pathway, tigecycline induced apoptosis. Furthermore, tigecycline were cytotoxic to both resistant cells. That means tigecycline could be used as the new strategy for treating the recurrent and resistant leukemia. ABT737 and BAY11-7821 also could be new agents, however, they have not been approved for the clinical use. Interestingly, ABT737 showed collateral sensitivity to the resistant cells. Tigecycline mechanically has both effects on leukemic cells. In short, tigecycline might give a marked improvement in anti-leukemic chemotherapy.
Session topic: Publication Only
Type: Publication Only
Background
Novel tetracycline derivative tigecycline has an antibiotic effect against a wide range of microorganisms including an activity in multi-resistant drug infections and modulates the production of cytokines and chemokines. FDA approved it for the treatment of complicated infections in 2005. Besides it might affect hematological malignancies. Skrtic et al. reported tigecycline’s anti-leukemia activity in TEX cell lines and M9-ENL-1 cell lines. The phase I clinical trial has undergone in USA.
Aims
We examined the anti-leukemic effect and elucidated the mechanism of tigecycline. Furthermore, we confirmed tigecycline is also effective on cytarabine resistant (AraC-R) HL60 cell line and clofarabine resistant (CAFdA-R) HL60 cell line. Two cell lines showed 200-fold and 100-fold resistant to parental HL60 cells, respectively.
Methods
To assess the effects of tigecycline on cell viability in acute myeloid leukemia HL60 cell line, acute lymphocytic leukemia CEM cell line, multiple myeloma U266 cell line and chronic myeloid leukemia K562 cell line, XTT assay was performed. Annexin-PI assay, sub G1 analysis in cell cycle were performed using by flowcytometry in HL60 and CEM cell line. Apoptosis related protein (caspase 3, 8, 9, Bax, Bad, Bclxl, Bcl2) and the phosphorylation of IKKα/β, NFκB were detected by Western Blotting. The effectiveness of tigecycline on AraC-R HL60 cell line and CAFdA-R HL60 cell line, those have been established in our laboratory, were evaluated. Sensitivities on Bcl2 inhibitor (ABT737) and NFkB inhibitor (BAY11-7821) in HL60, CEM, AraC-R HL60 and CAFdA-R HL60 cells, were also examined.
Results
1) The cell viability was measured at 72 hours by XTT assay. The values of Inhibition Concentration 50% (IC50) of tigecycline in HL60, CEM, K562 and U266 cells were 5.0μM, 2.5μM, 2.7μM and 21.2μM respectively. It was shown that tigecycline was cytotoxic to leukemic cells. 2) Tigecycline induced apoptosis in a dose- and time- dependent manner. In addition, sub G1 analysis showed that tigecycline suppressed S phase in cell cycle. 3) Tigecycline suppressed the expression Bcl2 in a time-dependent manner and inhibited the phosphorylation of NFκB in HL60 cells and CEM cells. Tigecycline activated caspase3, 9. 4) We examined the effectiveness of tigecycline on AraC-R HL60 cells and CAFdA-R HL60 cells. The values of IC50 of tigecycline were 6.7μM (AraC-R) and 2.0μM (CAFdA-R). ABT737 and BAY11-7821 displayed more effective on resistant cells compared to HL60 parental cells. The values of IC50 of ABT737 were 881nM (Parental), 73.3nM (AraC-R), 68.4nM (CAFdA-R). Those of BAY11-7821 were 2.4μM (Parental), 1.1μM (AraC-R), 1.5μM (CAFdA-R).
Summary
Tigecycline showed an anti-leukemic effect on HL60, CEM and K562 cell lines. Tigecycline is presently used for the treatment of certain infections. Pharmacological concentration of tigecycline is useful for treatment of cancer and leukemia. Targeting caspase3, 9 pathway and NFκB pathway, tigecycline induced apoptosis. Furthermore, tigecycline were cytotoxic to both resistant cells. That means tigecycline could be used as the new strategy for treating the recurrent and resistant leukemia. ABT737 and BAY11-7821 also could be new agents, however, they have not been approved for the clinical use. Interestingly, ABT737 showed collateral sensitivity to the resistant cells. Tigecycline mechanically has both effects on leukemic cells. In short, tigecycline might give a marked improvement in anti-leukemic chemotherapy.
Session topic: Publication Only
Abstract: PB1633
Type: Publication Only
Background
Novel tetracycline derivative tigecycline has an antibiotic effect against a wide range of microorganisms including an activity in multi-resistant drug infections and modulates the production of cytokines and chemokines. FDA approved it for the treatment of complicated infections in 2005. Besides it might affect hematological malignancies. Skrtic et al. reported tigecycline’s anti-leukemia activity in TEX cell lines and M9-ENL-1 cell lines. The phase I clinical trial has undergone in USA.
Aims
We examined the anti-leukemic effect and elucidated the mechanism of tigecycline. Furthermore, we confirmed tigecycline is also effective on cytarabine resistant (AraC-R) HL60 cell line and clofarabine resistant (CAFdA-R) HL60 cell line. Two cell lines showed 200-fold and 100-fold resistant to parental HL60 cells, respectively.
Methods
To assess the effects of tigecycline on cell viability in acute myeloid leukemia HL60 cell line, acute lymphocytic leukemia CEM cell line, multiple myeloma U266 cell line and chronic myeloid leukemia K562 cell line, XTT assay was performed. Annexin-PI assay, sub G1 analysis in cell cycle were performed using by flowcytometry in HL60 and CEM cell line. Apoptosis related protein (caspase 3, 8, 9, Bax, Bad, Bclxl, Bcl2) and the phosphorylation of IKKα/β, NFκB were detected by Western Blotting. The effectiveness of tigecycline on AraC-R HL60 cell line and CAFdA-R HL60 cell line, those have been established in our laboratory, were evaluated. Sensitivities on Bcl2 inhibitor (ABT737) and NFkB inhibitor (BAY11-7821) in HL60, CEM, AraC-R HL60 and CAFdA-R HL60 cells, were also examined.
Results
1) The cell viability was measured at 72 hours by XTT assay. The values of Inhibition Concentration 50% (IC50) of tigecycline in HL60, CEM, K562 and U266 cells were 5.0μM, 2.5μM, 2.7μM and 21.2μM respectively. It was shown that tigecycline was cytotoxic to leukemic cells. 2) Tigecycline induced apoptosis in a dose- and time- dependent manner. In addition, sub G1 analysis showed that tigecycline suppressed S phase in cell cycle. 3) Tigecycline suppressed the expression Bcl2 in a time-dependent manner and inhibited the phosphorylation of NFκB in HL60 cells and CEM cells. Tigecycline activated caspase3, 9. 4) We examined the effectiveness of tigecycline on AraC-R HL60 cells and CAFdA-R HL60 cells. The values of IC50 of tigecycline were 6.7μM (AraC-R) and 2.0μM (CAFdA-R). ABT737 and BAY11-7821 displayed more effective on resistant cells compared to HL60 parental cells. The values of IC50 of ABT737 were 881nM (Parental), 73.3nM (AraC-R), 68.4nM (CAFdA-R). Those of BAY11-7821 were 2.4μM (Parental), 1.1μM (AraC-R), 1.5μM (CAFdA-R).
Summary
Tigecycline showed an anti-leukemic effect on HL60, CEM and K562 cell lines. Tigecycline is presently used for the treatment of certain infections. Pharmacological concentration of tigecycline is useful for treatment of cancer and leukemia. Targeting caspase3, 9 pathway and NFκB pathway, tigecycline induced apoptosis. Furthermore, tigecycline were cytotoxic to both resistant cells. That means tigecycline could be used as the new strategy for treating the recurrent and resistant leukemia. ABT737 and BAY11-7821 also could be new agents, however, they have not been approved for the clinical use. Interestingly, ABT737 showed collateral sensitivity to the resistant cells. Tigecycline mechanically has both effects on leukemic cells. In short, tigecycline might give a marked improvement in anti-leukemic chemotherapy.
Session topic: Publication Only
Type: Publication Only
Background
Novel tetracycline derivative tigecycline has an antibiotic effect against a wide range of microorganisms including an activity in multi-resistant drug infections and modulates the production of cytokines and chemokines. FDA approved it for the treatment of complicated infections in 2005. Besides it might affect hematological malignancies. Skrtic et al. reported tigecycline’s anti-leukemia activity in TEX cell lines and M9-ENL-1 cell lines. The phase I clinical trial has undergone in USA.
Aims
We examined the anti-leukemic effect and elucidated the mechanism of tigecycline. Furthermore, we confirmed tigecycline is also effective on cytarabine resistant (AraC-R) HL60 cell line and clofarabine resistant (CAFdA-R) HL60 cell line. Two cell lines showed 200-fold and 100-fold resistant to parental HL60 cells, respectively.
Methods
To assess the effects of tigecycline on cell viability in acute myeloid leukemia HL60 cell line, acute lymphocytic leukemia CEM cell line, multiple myeloma U266 cell line and chronic myeloid leukemia K562 cell line, XTT assay was performed. Annexin-PI assay, sub G1 analysis in cell cycle were performed using by flowcytometry in HL60 and CEM cell line. Apoptosis related protein (caspase 3, 8, 9, Bax, Bad, Bclxl, Bcl2) and the phosphorylation of IKKα/β, NFκB were detected by Western Blotting. The effectiveness of tigecycline on AraC-R HL60 cell line and CAFdA-R HL60 cell line, those have been established in our laboratory, were evaluated. Sensitivities on Bcl2 inhibitor (ABT737) and NFkB inhibitor (BAY11-7821) in HL60, CEM, AraC-R HL60 and CAFdA-R HL60 cells, were also examined.
Results
1) The cell viability was measured at 72 hours by XTT assay. The values of Inhibition Concentration 50% (IC50) of tigecycline in HL60, CEM, K562 and U266 cells were 5.0μM, 2.5μM, 2.7μM and 21.2μM respectively. It was shown that tigecycline was cytotoxic to leukemic cells. 2) Tigecycline induced apoptosis in a dose- and time- dependent manner. In addition, sub G1 analysis showed that tigecycline suppressed S phase in cell cycle. 3) Tigecycline suppressed the expression Bcl2 in a time-dependent manner and inhibited the phosphorylation of NFκB in HL60 cells and CEM cells. Tigecycline activated caspase3, 9. 4) We examined the effectiveness of tigecycline on AraC-R HL60 cells and CAFdA-R HL60 cells. The values of IC50 of tigecycline were 6.7μM (AraC-R) and 2.0μM (CAFdA-R). ABT737 and BAY11-7821 displayed more effective on resistant cells compared to HL60 parental cells. The values of IC50 of ABT737 were 881nM (Parental), 73.3nM (AraC-R), 68.4nM (CAFdA-R). Those of BAY11-7821 were 2.4μM (Parental), 1.1μM (AraC-R), 1.5μM (CAFdA-R).
Summary
Tigecycline showed an anti-leukemic effect on HL60, CEM and K562 cell lines. Tigecycline is presently used for the treatment of certain infections. Pharmacological concentration of tigecycline is useful for treatment of cancer and leukemia. Targeting caspase3, 9 pathway and NFκB pathway, tigecycline induced apoptosis. Furthermore, tigecycline were cytotoxic to both resistant cells. That means tigecycline could be used as the new strategy for treating the recurrent and resistant leukemia. ABT737 and BAY11-7821 also could be new agents, however, they have not been approved for the clinical use. Interestingly, ABT737 showed collateral sensitivity to the resistant cells. Tigecycline mechanically has both effects on leukemic cells. In short, tigecycline might give a marked improvement in anti-leukemic chemotherapy.
Session topic: Publication Only
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