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http://dx.doi.org/10.18419/opus-8262
Autor(en): | Möller, Yvonne Siegemund, Martin Beyes, Sven Herr, Ricarda Lecis, Daniele Delia, Domenico Kontermann, Roland Brummer, Tilman Pfizenmaier, Klaus Olayioye, Monilola A. |
Titel: | EGFR-targeted TRAIL and a Smac mimetic synergize to overcome apoptosis resistance in KRAS mutant colorectal cancer cells |
Erscheinungsdatum: | 2014 |
Dokumentart: | Zeitschriftenartikel |
Erschienen in: | PLoS one 9 (2014), issue 9, e107165. URL http://dx.doi.org./10.1371/journal.pone.0107165 |
URI: | http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-95663 http://elib.uni-stuttgart.de/handle/11682/8279 http://dx.doi.org/10.18419/opus-8262 |
Zusammenfassung: | TRAIL is a death receptor ligand that induces cell death preferentially in tumor cells. Recombinant soluble TRAIL, however, performs poorly as an anti-cancer therapeutic because oligomerization is required for potent biological activity. We previously generated a diabody format of tumor-targeted TRAIL termed Db<sub>αEGFR</sub>-scTRAIL, comprising single-stranded TRAIL molecules (scTRAIL) and the variable domains of a humanized variant of the EGFR blocking antibody Cetuximab. Here we define the bioactivity of Db<sub>αEGFR</sub>-scTRAIL with regard to both EGFR inhibition and TRAIL receptor activation in 3D cultures of Caco-2 colorectal cancer cells, which express wild-type K-Ras. Compared with conventional 2D cultures, Caco-2 cells displayed strongly enhanced sensitivity toward Db<sub>αEGFR</sub>-scTRAIL in these 3D cultures. We show that the antibody moiety of Db<sub>αEGFR</sub>-scTRAIL not only efficiently competed with ligand-induced EGFR function, but also determined the apoptotic response by specifically directing Db<sub>αEGFR</sub>-scTRAIL to EGFR-positive cells. To address how aberrantly activated K-Ras, which leads to Cetuximab resistance, affects Db<sub>αEGFR</sub>-scTRAIL sensitivity, we generated stable Caco-2tet cells inducibly expressing oncogenic K-Ras<sup>G12V</sup>. In the presence of doxycycline, these cells showed increased resistance to Db<sub>αEGFR</sub>-scTRAIL, associated with the elevated expression of the anti-apoptotic proteins cIAP2, Bcl-xL and Flip<sub>S</sub>. Co-treatment of cells with the Smac mimetic SM83 restored the Db<sub>αEGFR</sub>-scTRAIL-induced apoptotic response. Importantly, this synergy between Db<sub>αEGFR</sub>-scTRAIL and SM83 also translated to 3D cultures of oncogenic K-Ras expressing HCT-116 and LoVo colorectal cancer cells. Our findings thus support the notion that Db<sub>αEGFR</sub>-scTRAIL therapy in combination with apoptosis-sensitizing agents may be promising for the treatment of EGFR-positive colorectal cancers, independently of their KRAS status. |
Enthalten in den Sammlungen: | 15 Fakultätsübergreifend / Sonstige Einrichtung |
Dateien zu dieser Ressource:
Datei | Beschreibung | Größe | Format | |
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001_journal.pone.0107165.pdf | Article | 1,35 MB | Adobe PDF | Öffnen/Anzeigen |
01_Figure_S1.pdf | Supplementary figure 1 | 214,21 kB | Adobe PDF | Öffnen/Anzeigen |
02_Figure_S2.pdf | Supplementary figure 2 | 134,92 kB | Adobe PDF | Öffnen/Anzeigen |
03_Figure_S3.pdf | Supplementary figure 3 | 95,52 kB | Adobe PDF | Öffnen/Anzeigen |
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