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Galanin and its receptors,  GALR 1 and  GALR 2, are known tumor suppressors and potential therapeutic targets in head and neck squamous cell carcinoma ( HNSCC ). Previously, we demonstrated that, in  GALR 1‐expressing  HNSCC  cells, the addition of galanin suppressed tumor proliferation via upregulation of  ERK 1/2 and cyclin‐dependent kinase inhibitors, whereas, in  GALR 2‐expressing cells, the addition of galanin not only suppressed proliferation, but also induced apoptosis. In this study, we first transduced  HEp ‐2 and  KB  cell lines using a recombinant adeno‐associated virus ( rAAV )‐green fluorescent protein ( GFP ) vector and confirmed a high  GFP  expression rate (>90%) in both cell lines at the standard vector dose. Next, we demonstrated that  GALR 2 expression in the presence of galanin suppressed cell viability to 40–60% after 72 h in both cell lines. Additionally, the annexin V‐positive rate and sub‐G0/G1 phase population were significantly elevated in  HEp ‐2 cells (mock  vs   GALR 2: 12.3  vs  25.0% ( P  < 0.01) and 9.1  vs  32.0% ( P  < 0.05), respectively) after 48 h. These changes were also observed in  KB  cells, although to a lesser extent. Furthermore, in  HEp ‐2 cells,  GALR 2‐mediated apoptosis was caspase‐independent, involving downregulation of  ERK 1/2, followed by induction of the pro‐apoptotic Bcl‐2 protein, Bim. These results illustrate that transient  GALR 2 expression in the presence of galanin induces apoptosis via diverse pathways and serves as a platform for suicide gene therapy against  HNSCC .

Novel anti‐tumor mechanism of galanin receptor type 2 in head and neck squamous cell carcinoma cells