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Receptor activation and inhibition in cellular response to chemotherapeutic combinational mimicries: the concept of divergent targeting

  • Laboratory Investigation - Human/Animal Tissue
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Abstract

The antiproliferative effect of tandem somatostatin receptor (SSTR) activation, epidermal growth factor receptor (EGFR) inhibition, and induction of DNA damage was analyzed using octreotide (OCT), a SSTR agonist, the clinical DNA methylating agent temozolomide (TMZ), Iressa, an EGFR inhibitor, and dual EGFR-DNA targeting agents termed “combi-molecules”. Using SSTR-expressing glioma cells harbouring low levels of EGFR (U87MG) or transfected to overexpress EGFR (U87/EGFR) or a variant (U87/EGFRvIII), we showed that Iressa, alone or in combination with the DNA damaging agent TMZ, and combi-molecules RA2 and RA5 inhibited EGF-induced phosphorylation of EGFR in U87MG and more moderately in U87/EGFR and U87/EGFRvIII transfected cells. This translated into equivalent levels of Erk 1/2 inhibition. Activation of SSTRs with OCT did not modulate the effects of the various treatments on Erk 1/2 phosphorylation. Likewise, SSTR activation did not alter TMZ- or DNA-damaging combi-molecules, RA2 and RA5, induced p53 activation nor upregulation. However, SSTR activation significantly shifted TMZ-, RA2- and RA5-induced cell-cycle arrest to earlier phases (i.e., G2/M to late S, late S to S, S to G1). Further analysis showed that apoptosis was not induced. This was in agreement with the fact that p53 activation did not induce Bax upregulation nor did EGFR inhibition promote Bad dephosphorylation. Moreover, enhancement of survivin, an anti-apoptotic protein, expression was observed. The results in toto suggest that the combination of SSTR activation with EGFR inhibition and DNA damage affects cell-cycle progression but a disconnection between the targeted signalling pathways in these brain tumour cells precludes synergistic cell-killing by the triple growth inhibitory events.

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Abbreviations

ATM:

Ataxia-telangiectasia

ATR:

ATM and Rad-3-related

BER:

Base excision repair

DMSO:

Dimethyl sulfoxide

EGF:

Epidermal growth factor

ErbBs:

Epidermal growth factor receptors

GPCR:

G protein-coupled receptors

MAPK:

Mitogen-activated protein kinase

NER:

Nucleotide excision repair

OCT:

Octreotide

PBST:

0.1% Tween 20 (v/v) in phosphate-buffered saline

PGT:

Poly(L-glutamic acid–L-tyrosine 4:1)

PI:

Propidium iodide

PVDF:

Poly(vinylidene difluoride)

RTK:

Receptor tyrosine kinase

SST:

Somatostatin

SSTR:

Somatostatin receptor

TK:

Tyrosine kinase

TKI:

Tyrosine kinase inhibitor

TMB:

Tetramethylbenzidine peroxidase substrate

TMZ:

Temozolomide

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Acknowledgements

We would like to acknowledge the Canadian Institute for Health Research (CIHR) and the Brain Tumour Foundation of Canada for financial support. HW was supported by a McGill University Health Centre Research Institute Studentship.

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  1. Cancer Drug Research Laboratory, Department of Medicine, McGill University Health Centre/Royal Victoria Hospital, 687 Pine Ave West, Room M7.15, Montreal, QC, H3A 1A1, Canada

    H. L. Watt, Z. Rachid & B. J. Jean-Claude

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  1. H. L. Watt
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  2. Z. Rachid
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  3. B. J. Jean-Claude
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Correspondence to B. J. Jean-Claude.

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Watt, H.L., Rachid, Z. & Jean-Claude, B.J. Receptor activation and inhibition in cellular response to chemotherapeutic combinational mimicries: the concept of divergent targeting. J Neurooncol 100, 345–361 (2010). https://doi.org/10.1007/s11060-010-0196-7

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