| Autor: |
Todorova MI; Cancer Drug Research Laboratory, Department of Medicine, Division of Medical Oncology, McGill University/Royal Victoria Hospital, 687 Pine Avenue West, Montreal, Quebec, Canada H3A 1A1., Larroque AL, Dauphin-Pierre S, Fang YQ, Jean-Claude BJ |
| Jazyk: |
angličtina |
| Zdroj: |
Molecular cancer therapeutics [Mol Cancer Ther] 2010 Apr; Vol. 9 (4), pp. 869-82. Date of Electronic Publication: 2010 Mar 30. |
| DOI: |
10.1158/1535-7163.MCT-09-0673 |
| Abstrakt: |
To monitor the subcellular distribution of mixed epidermal growth factor (EGF) receptor (EGFR)-DNA targeting drugs termed combi-molecules, we designed AL237, a fluorescent prototype, to degrade into a green fluorescent DNA damaging species and FD105, a blue fluorescent EGFR inhibitor. Here we showed that AL237 damaged DNA in the 12.5 to 50 mumol/L range. Despite its size, it blocked EGFR phosphorylation in an enzyme assay (IC(50) = 0.27 mumol/L) and in MDA-MB468 breast cancer cells in the same concentration range as for DNA damage. This translated into inhibition of extracellular signal-regulated kinase 1/2 or BAD phosphorylation and downregulation of DNA repair proteins (XRCC1, ERCC1). Having shown that AL237 was a balanced EGFR-DNA targeting molecule, it was used as an imaging probe to show that (a) green and blue colors were primarily colocalized in the perinuclear and partially in the nucleus in EGFR- or ErbB2-expressing cells, (b) the blue fluorescence associated with FD105, but not the green, was colocalized with anti-EGFR red-labeled antibody, (c) the green fluorescence of nuclei was significantly more intense in NIH 3T3 cells expressing EGFR or ErbB2 than in their wild-type counterparts (P < 0.05). Similarly, the growth inhibitory potency of AL237 was selectively stronger in the transfectants. In summary, the results suggest that AL237 diffuses into the cells and localizes abundantly in the perinuclear region and partially in the nucleus where it degrades into EGFR and DNA targeting species. This bystander-like effect translates into high levels of DNA damage in the nucleus. Sufficient quinazoline levels are released in the cells to block EGF-induced activation of downstream signaling. Mol Cancer Ther; 9(4); 869-82. (c)2010 AACR. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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