Multiple signalling events interact in cancer cells. Oncogenic Ras cooperates with Egfr, which cannot be explained by the canonical signalling paradigm.

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In turn, Egfr cooperates with Hedgehog signalling. How oncogenic Ras elicits and integrates Egfr and Hedgehog signals to drive overgrowth remains unclear.

Using a Drosophila tumour model, we show that Egfr cooperates with oncogenic Ras via Arf6, which functions as a novel regulator of Hh signalling. Oncogenic Ras induces the expression of Egfr ligands. Egfr then signals through Arf6, which regulates Hh transport to promote Hh signalling.

Blocking any step of this signalling cascade inhibits Hh signalling and correspondingly suppresses the growth of both, fly and human cancer cells harbouring oncogenic Ras mutations. These findings highlight a non-canonical Egfr signalling mechanism, centered on Arf6 as a novel regulator of Hh signalling. This explains both, the puzzling requirement of Egfr in oncogenic Ras-mediated overgrowth and the cooperation between Egfr and Hedgehog. Activating mutations of the Ras gene are highly prevalent in human cancers and give rise to some of the most aggressive tumours. The molecular mechanisms governing oncogenic Ras-driven cancers are complex and involve interacting signalling pathways. Paradoxically, oncogenic Ras cooperates with Egfr in cancers. Torrent crack anno 1701 serial. EGFR ligands bind to and activate Egfr, which recruits docking proteins via its cytoplasmic domain.

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Docking proteins (such as the downstream of receptor kinases or drk) activate the guanine exchange factor Son of sevenless (Sos), which converts Ras from an inactive (GDP-bound) to an active (GTP-bound) state and leads to the MAPK signalling cascade and activation of downstream target molecules. It is surprising that the action of an activated downstream oncogenic component still requires its upstream receptor. On the other hand, Egfr has been shown to cooperate with Hedgehog (Hh) signalling, another oncogenic pathway, to drive basal cell carcinoma and melanomas. Hh signalling is initiated by the interaction of Hh protein with its receptor Patched. Endocytosis and intracellular transport of the receptor–ligand complex modulate Hh signalling levels.

How oncogenic Ras, Egfr and Hh signalling are integrated to concertedly drive tumour overgrowth remains unclear. Animal models expand our understanding of oncogenic Ras signalling. Using a fly tumour model of oncogenic Ras we have identified Egfr as a positive regulator of oncogenic Ras-mediated overgrowth. Our characterization of Egfr’s role in oncogenic Ras-mediated overgrowth led the finding that oncogenic Ras signalling stimulates the expression of the Egfr ligand spitz (spi) to recruit Egfr signalling and achieve tumour overgrowth. Egfr promotes tumour overgrowth independent of the canonical the Sos/Ras signalling, instead it acts via the ADP-Ribosylation Factor 6 (Arf6). Arf6 belongs to a family of highly conserved small Ras-related GTP-binding proteins and is largely known for its role in regulating endocytosis, vesicle transport and secretion,,,,,. We investigated a role for Arf6 in oncogenic Ras tumour overgrowth and found that Egfr promotes Arf6 to interact with Hh.

This interaction allows Arf6 to control Hh cellular trafficking and promote Hh signalling. Consistent with this, blocking Egfr or Arf6 suppresses Hh signalling and inhibits the growth of either fly or human cancer cells harbouring oncogenic Ras.

Altogether, our data delineate a non-canonical Egfr signalling mechanism in which Arf6 acts as a novel regulator of Hh signalling. This explains the puzzling requirement of Egfr in oncogenic Ras-mediated overgrowth and the oncogenic cooperation between Egfr and Hh signalling. Egfr − suppresses Ras V12 tumours by inhibiting cell proliferation Mosaic expression of oncogenic Ras ( Ras V12) gives rise to hyperplastic tumours in Drosophila tissues.

These tumours can be GFP-labelled and a measure of the overgrowth phenotype can be readily obtained by examining the size and the fluorescence intensity of clones in dissected eye-antenna imaginal discs from third-instar animals, (). We searched for mutations that suppress Ras V12 tumour overgrowth and identified a null Egfr mutation Egfr Co (ref. ), hereafter referred to as Egfr _. We generated clones of cells harbouring the Egfr _ mutation or expressing Ras V12 in the presence of the Egfr _ mutation and scored clones size. Consistent with EGFR’s known role in controlling cell survival and growth, Egfr _ mutant cells yielded small clones (). We found that Egfr _ suppressed Ras V12 tumour overgrowth (; quantified in ). A dominant-negative version of Egfr (lacking its cytoplasmic tail) produced a similar effect ().