Limits...
The effect of the dual Src/Abl kinase inhibitor AZD0530 on Philadelphia positive leukaemia cell lines.

Gwanmesia PM, Romanski A, Schwarz K, Bacic B, Ruthardt M, Ottmann OG - BMC Cancer (2009)

Bottom Line: An ongoing transphosphorylation was demonstrated between SFKs and Bcr-Abl.AZD0530 significantly down-regulated the activation of survival signalling pathways in Ph+ cells, resistant or sensitive to Imatinib, with the exception of the RTSupB15.Our results indicate that AZD0530 targets both Src and Bcr-Abl kinase activity and reduces the leukaemic maintenance by Bcr-Abl.

View Article: PubMed Central - HTML - PubMed

Affiliation: Med, Klinik II/Abt, Hämatologie, Johann Wolfgang Goethe-Universität, 60590 Frankfurt, Germany. Mambou@uni-heidelberg.de

ABSTRACT

Background: Imatinib mesylate, a selective inhibitor of Abl tyrosine kinase, is efficacious in treating chronic myeloid leukaemia (CML) and Ph+ acute lymphoblastic leukaemia (ALL). However, most advanced-phase CML and Ph+ ALL patients relapse on Imatinib therapy. Several mechanisms of refractoriness have been reported, including the activation of the Src-family kinases (SFK). Here, we investigated the biological effect of the new specific dual Src/Abl kinase inhibitor AZD0530 on Ph+ leukaemic cells.

Methods: Cell lines used included BV173 (CML in myeloid blast crisis), SEM t(4;11), Ba/F3 (IL-3 dependent murine pro B), p185Bcr-Abl infected Ba/F3 cells, p185Bcr-Abl mutant infected Ba/F3 cells, SupB15 (Ph+ ALL) and Imatinib resistant SupB15 (RTSupB15) (Ph+ ALL) cells. Cells were exposed to AZD0530 and Imatinib. Cell proliferation, apoptosis, survival and signalling pathways were assessed by dye exclusion, flow cytometry and Western blotting respectively.

Results: AZD0530 specifically inhibited the growth of, and induced apoptosis in CML and Ph+ ALL cells in a dose dependent manner, but showed only marginal effects on Ph- ALL cells. Resistance to Imatinib due to the mutation Y253F in p185Bcr-Abl was overcome by AZD0530. Combination of AZD0530 and Imatinib showed an additive inhibitory effect on the proliferation of CML BV173 cells but not on Ph+ ALL SupB15 cells. An ongoing transphosphorylation was demonstrated between SFKs and Bcr-Abl. AZD0530 significantly down-regulated the activation of survival signalling pathways in Ph+ cells, resistant or sensitive to Imatinib, with the exception of the RTSupB15.

Conclusion: Our results indicate that AZD0530 targets both Src and Bcr-Abl kinase activity and reduces the leukaemic maintenance by Bcr-Abl.

Show MeSH

Related in: MedlinePlus

Effects of AZD0530 on proliferation. A AZD0530 specifically blocks the proliferation of Ph+ BV173 cells in a dose dependent manner, while the proliferation of the Ph- SEM cells is not markedly affected. B AZD0530 inhibited proliferation of WTSupB15, in a dose-dependent way. No effect was observed on the proliferation of RTSupB15 upon treatment with AZD0530. C AZD0530 inhibited the proliferation of Ba/F3 cells expressing WTp185Bcr-Abl and the Bcr-Abl Mut Y253F. Ba/F3 cells expressing the Bcr-Abl Mut E255K were less sensitive to AZD0530 as compared to Mut Y253F and cells expressing Bcr-Abl Mut T315I were stable in the presence of AZD0530. All cell lines were treated with the indicated concentrations of AZD0530 for three days and proliferation was determined by trypan blue exclusion of viable cells. Results are the mean values of 3 independent experiments carried out in duplicates +/- S.D.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2654659&req=5

Figure 1: Effects of AZD0530 on proliferation. A AZD0530 specifically blocks the proliferation of Ph+ BV173 cells in a dose dependent manner, while the proliferation of the Ph- SEM cells is not markedly affected. B AZD0530 inhibited proliferation of WTSupB15, in a dose-dependent way. No effect was observed on the proliferation of RTSupB15 upon treatment with AZD0530. C AZD0530 inhibited the proliferation of Ba/F3 cells expressing WTp185Bcr-Abl and the Bcr-Abl Mut Y253F. Ba/F3 cells expressing the Bcr-Abl Mut E255K were less sensitive to AZD0530 as compared to Mut Y253F and cells expressing Bcr-Abl Mut T315I were stable in the presence of AZD0530. All cell lines were treated with the indicated concentrations of AZD0530 for three days and proliferation was determined by trypan blue exclusion of viable cells. Results are the mean values of 3 independent experiments carried out in duplicates +/- S.D.

Mentions: To determine the effect and specificity of AZD0530 on Src and Bcr-Abl mediated growth inhibition of Ph+ cells, the CML blast cell line BV173, was treated with various concentrations of AZD0530, and cell proliferation was measured by trypan blue exclusion of viable cells. As control for specificity the Ph- ALL cell line SEM, was used. Figure 1A shows that incubation with AZD0530, resulted in a dose-dependent decrease in cell proliferation of BV173 cells in contrast to the SEM cells over a three-day incubation period. In the BV173 cells, growth inhibition could be observed as from 0,5 μM AZD0530 when compared to DMSO treated cells. Proliferation in the SEM cells was not affected in the presence of 5 μM AZD0530 when compared to control cells.


The effect of the dual Src/Abl kinase inhibitor AZD0530 on Philadelphia positive leukaemia cell lines.

Gwanmesia PM, Romanski A, Schwarz K, Bacic B, Ruthardt M, Ottmann OG - BMC Cancer (2009)

Effects of AZD0530 on proliferation. A AZD0530 specifically blocks the proliferation of Ph+ BV173 cells in a dose dependent manner, while the proliferation of the Ph- SEM cells is not markedly affected. B AZD0530 inhibited proliferation of WTSupB15, in a dose-dependent way. No effect was observed on the proliferation of RTSupB15 upon treatment with AZD0530. C AZD0530 inhibited the proliferation of Ba/F3 cells expressing WTp185Bcr-Abl and the Bcr-Abl Mut Y253F. Ba/F3 cells expressing the Bcr-Abl Mut E255K were less sensitive to AZD0530 as compared to Mut Y253F and cells expressing Bcr-Abl Mut T315I were stable in the presence of AZD0530. All cell lines were treated with the indicated concentrations of AZD0530 for three days and proliferation was determined by trypan blue exclusion of viable cells. Results are the mean values of 3 independent experiments carried out in duplicates +/- S.D.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654659&req=5

Figure 1: Effects of AZD0530 on proliferation. A AZD0530 specifically blocks the proliferation of Ph+ BV173 cells in a dose dependent manner, while the proliferation of the Ph- SEM cells is not markedly affected. B AZD0530 inhibited proliferation of WTSupB15, in a dose-dependent way. No effect was observed on the proliferation of RTSupB15 upon treatment with AZD0530. C AZD0530 inhibited the proliferation of Ba/F3 cells expressing WTp185Bcr-Abl and the Bcr-Abl Mut Y253F. Ba/F3 cells expressing the Bcr-Abl Mut E255K were less sensitive to AZD0530 as compared to Mut Y253F and cells expressing Bcr-Abl Mut T315I were stable in the presence of AZD0530. All cell lines were treated with the indicated concentrations of AZD0530 for three days and proliferation was determined by trypan blue exclusion of viable cells. Results are the mean values of 3 independent experiments carried out in duplicates +/- S.D.
Mentions: To determine the effect and specificity of AZD0530 on Src and Bcr-Abl mediated growth inhibition of Ph+ cells, the CML blast cell line BV173, was treated with various concentrations of AZD0530, and cell proliferation was measured by trypan blue exclusion of viable cells. As control for specificity the Ph- ALL cell line SEM, was used. Figure 1A shows that incubation with AZD0530, resulted in a dose-dependent decrease in cell proliferation of BV173 cells in contrast to the SEM cells over a three-day incubation period. In the BV173 cells, growth inhibition could be observed as from 0,5 μM AZD0530 when compared to DMSO treated cells. Proliferation in the SEM cells was not affected in the presence of 5 μM AZD0530 when compared to control cells.

Bottom Line: An ongoing transphosphorylation was demonstrated between SFKs and Bcr-Abl.AZD0530 significantly down-regulated the activation of survival signalling pathways in Ph+ cells, resistant or sensitive to Imatinib, with the exception of the RTSupB15.Our results indicate that AZD0530 targets both Src and Bcr-Abl kinase activity and reduces the leukaemic maintenance by Bcr-Abl.

View Article: PubMed Central - HTML - PubMed

Affiliation: Med, Klinik II/Abt, Hämatologie, Johann Wolfgang Goethe-Universität, 60590 Frankfurt, Germany. Mambou@uni-heidelberg.de

ABSTRACT

Background: Imatinib mesylate, a selective inhibitor of Abl tyrosine kinase, is efficacious in treating chronic myeloid leukaemia (CML) and Ph+ acute lymphoblastic leukaemia (ALL). However, most advanced-phase CML and Ph+ ALL patients relapse on Imatinib therapy. Several mechanisms of refractoriness have been reported, including the activation of the Src-family kinases (SFK). Here, we investigated the biological effect of the new specific dual Src/Abl kinase inhibitor AZD0530 on Ph+ leukaemic cells.

Methods: Cell lines used included BV173 (CML in myeloid blast crisis), SEM t(4;11), Ba/F3 (IL-3 dependent murine pro B), p185Bcr-Abl infected Ba/F3 cells, p185Bcr-Abl mutant infected Ba/F3 cells, SupB15 (Ph+ ALL) and Imatinib resistant SupB15 (RTSupB15) (Ph+ ALL) cells. Cells were exposed to AZD0530 and Imatinib. Cell proliferation, apoptosis, survival and signalling pathways were assessed by dye exclusion, flow cytometry and Western blotting respectively.

Results: AZD0530 specifically inhibited the growth of, and induced apoptosis in CML and Ph+ ALL cells in a dose dependent manner, but showed only marginal effects on Ph- ALL cells. Resistance to Imatinib due to the mutation Y253F in p185Bcr-Abl was overcome by AZD0530. Combination of AZD0530 and Imatinib showed an additive inhibitory effect on the proliferation of CML BV173 cells but not on Ph+ ALL SupB15 cells. An ongoing transphosphorylation was demonstrated between SFKs and Bcr-Abl. AZD0530 significantly down-regulated the activation of survival signalling pathways in Ph+ cells, resistant or sensitive to Imatinib, with the exception of the RTSupB15.

Conclusion: Our results indicate that AZD0530 targets both Src and Bcr-Abl kinase activity and reduces the leukaemic maintenance by Bcr-Abl.

Show MeSH
Related in: MedlinePlus