To define the mechanisms regulating cell survival and growth of HNSCC, HSC–3 cells were subjected to cell culture in 3 dimensions (3D culture) as multicellular aggregates (MCA) also called spheroids. ErbB3 expression was significantly increased following MCA formation compared with monolayer (ML) culture at 24 h (not shown here).To determine the importance of elevated ErbB3 in downstream signalling, MCA were treated with the ErbB3 ligand, HRG–β1 or with EGF as a reference control. HRG–β1 induced strong p–ErbB3 compared with the non–treated control (not shown). In addition, HRG–β1 treatment also increased the p–ErbB2 level and led to significant induction of p–AKT. Like HRG–β1, EGF treatment also induced p–ErbB3, albeit at lower levels. As expected, EGF treatment led to increased induction of p–ErbB2, p–EGFR and p–AKT levels.The authors next tested if elevated ErbB3 level may modulate cell survival and growth.

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