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Effect of Aplidin in acute lymphoblastic leukaemia cells.

Erba E, Serafini M, Gaipa G, Tognon G, Marchini S, Celli N, Rotilio D, Broggini M, Jimeno J, Faircloth GT, Biondi A, D'Incalci M - Br. J. Cancer (2003)

Bottom Line: Only in ALL-PO 20 nM Aplidin treatment caused a block of vascular endothelial growth factor (VEGF) secretion and downregulation of VEGF-mRNA, but Aplidin cytotoxicity does not seem to be related to VEGF inhibition since the sensitivity of ALL-PO cells to Aplidin is comparable to that observed for the other cells used.Cells from children with genetic abnormalities such as t(9;22) and t(4;11) translocations, associated with an inferior treatment outcome, were sensitive to Aplidin to the same extent as that observed in other BCP-ALL cases.Aplidin exerted a strong cell killing effect (>88%) against primary culture cells from five relapsed ALL cases, at concentrations much lower than those reported to be achieved in plasma of patients receiving Aplidin at recommended doses.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Istituto di Ricerche Farmacologiche 'Mario Negri', Via Eritrea, 62-20157 Milan, Italy. erba@marionegri.it

ABSTRACT
The cytotoxic effect of Aplidin was investigated on fresh leukaemia cells derived from children with B-cell-precursor (BCP) acute lymphoblastic leukaemia (ALL) by using stromal-layer culture system and on four cell lines, ALL-PO, Reh, ALL/MIK and TOM-1, derived from patients with ALL with different molecular genetic abnormalities. In ALL cell lines Aplidin was cytotoxic at nanomolar concentrations. In the ALL cell lines the drug-induced cell death was clearly related to the induction of apoptosis and appeared to be p53-independent. Only in ALL-PO 20 nM Aplidin treatment caused a block of vascular endothelial growth factor (VEGF) secretion and downregulation of VEGF-mRNA, but Aplidin cytotoxicity does not seem to be related to VEGF inhibition since the sensitivity of ALL-PO cells to Aplidin is comparable to that observed for the other cells used. Aplidin induced a G(1) and a G(2) M block in ALL cell lines. In patient-derived leukaemia cells, Aplidin induced a strong cytotoxicity evidenced in a stroma-supported immunocytometric assay. Cells from children with genetic abnormalities such as t(9;22) and t(4;11) translocations, associated with an inferior treatment outcome, were sensitive to Aplidin to the same extent as that observed in other BCP-ALL cases. Aplidin exerted a strong cell killing effect (>88%) against primary culture cells from five relapsed ALL cases, at concentrations much lower than those reported to be achieved in plasma of patients receiving Aplidin at recommended doses. Taken together these data suggest that Aplidin could be a new anticancer drug to be investigated in ALL patients resistant to available therapy.

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(A) Detection of apoptosis in ALL cells by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and the biparametric FSC/TdT-dUTP analysis were performed after 72 h after drug-washout. (B) Percentage of apoptotic cells evaluated by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and biparametric FSC/TdT-dUTP analysis were performed at different time intervals after drug-washout.
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fig4: (A) Detection of apoptosis in ALL cells by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and the biparametric FSC/TdT-dUTP analysis were performed after 72 h after drug-washout. (B) Percentage of apoptotic cells evaluated by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and biparametric FSC/TdT-dUTP analysis were performed at different time intervals after drug-washout.

Mentions: It has been reported that Aplidin acts in vitro by inducing apoptosis on different cells lines (Grubb et al, 1995; Erba et al, 2002). Aplidin induced apoptosis in all the cell lines used, as clearly seen by morphological examination by means of sulforhodamine/Dapi staining (data not shown). The level of apoptotic cells was different between the four cell lines as shown in Figure 4AFigure 4


Effect of Aplidin in acute lymphoblastic leukaemia cells.

Erba E, Serafini M, Gaipa G, Tognon G, Marchini S, Celli N, Rotilio D, Broggini M, Jimeno J, Faircloth GT, Biondi A, D'Incalci M - Br. J. Cancer (2003)

(A) Detection of apoptosis in ALL cells by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and the biparametric FSC/TdT-dUTP analysis were performed after 72 h after drug-washout. (B) Percentage of apoptotic cells evaluated by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and biparametric FSC/TdT-dUTP analysis were performed at different time intervals after drug-washout.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376915&req=5

fig4: (A) Detection of apoptosis in ALL cells by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and the biparametric FSC/TdT-dUTP analysis were performed after 72 h after drug-washout. (B) Percentage of apoptotic cells evaluated by TdT-dUTP flow cytometric analysis. Cells were treated with different concentrations of Aplidin and biparametric FSC/TdT-dUTP analysis were performed at different time intervals after drug-washout.
Mentions: It has been reported that Aplidin acts in vitro by inducing apoptosis on different cells lines (Grubb et al, 1995; Erba et al, 2002). Aplidin induced apoptosis in all the cell lines used, as clearly seen by morphological examination by means of sulforhodamine/Dapi staining (data not shown). The level of apoptotic cells was different between the four cell lines as shown in Figure 4AFigure 4

Bottom Line: Only in ALL-PO 20 nM Aplidin treatment caused a block of vascular endothelial growth factor (VEGF) secretion and downregulation of VEGF-mRNA, but Aplidin cytotoxicity does not seem to be related to VEGF inhibition since the sensitivity of ALL-PO cells to Aplidin is comparable to that observed for the other cells used.Cells from children with genetic abnormalities such as t(9;22) and t(4;11) translocations, associated with an inferior treatment outcome, were sensitive to Aplidin to the same extent as that observed in other BCP-ALL cases.Aplidin exerted a strong cell killing effect (>88%) against primary culture cells from five relapsed ALL cases, at concentrations much lower than those reported to be achieved in plasma of patients receiving Aplidin at recommended doses.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Istituto di Ricerche Farmacologiche 'Mario Negri', Via Eritrea, 62-20157 Milan, Italy. erba@marionegri.it

ABSTRACT
The cytotoxic effect of Aplidin was investigated on fresh leukaemia cells derived from children with B-cell-precursor (BCP) acute lymphoblastic leukaemia (ALL) by using stromal-layer culture system and on four cell lines, ALL-PO, Reh, ALL/MIK and TOM-1, derived from patients with ALL with different molecular genetic abnormalities. In ALL cell lines Aplidin was cytotoxic at nanomolar concentrations. In the ALL cell lines the drug-induced cell death was clearly related to the induction of apoptosis and appeared to be p53-independent. Only in ALL-PO 20 nM Aplidin treatment caused a block of vascular endothelial growth factor (VEGF) secretion and downregulation of VEGF-mRNA, but Aplidin cytotoxicity does not seem to be related to VEGF inhibition since the sensitivity of ALL-PO cells to Aplidin is comparable to that observed for the other cells used. Aplidin induced a G(1) and a G(2) M block in ALL cell lines. In patient-derived leukaemia cells, Aplidin induced a strong cytotoxicity evidenced in a stroma-supported immunocytometric assay. Cells from children with genetic abnormalities such as t(9;22) and t(4;11) translocations, associated with an inferior treatment outcome, were sensitive to Aplidin to the same extent as that observed in other BCP-ALL cases. Aplidin exerted a strong cell killing effect (>88%) against primary culture cells from five relapsed ALL cases, at concentrations much lower than those reported to be achieved in plasma of patients receiving Aplidin at recommended doses. Taken together these data suggest that Aplidin could be a new anticancer drug to be investigated in ALL patients resistant to available therapy.

Show MeSH
Related in: MedlinePlus