Preclinical study for Head and Neck Cancer using rAAV-GALR2

甘丙氨酸及其受体GALR1和GALR2是头颈部鳞状细胞癌（HNSCC）中已知的肿瘤抑制因子和潜在的治疗靶点。在这项研究中，我们首先使用重组腺相关病毒(rAAV)-绿色荧光蛋白（GFP）载体转导HEp-2和KB细胞系，并证实在这两种细胞系中GFP的高表达率（bbb90 %）。接下来，我们证明了GALR2在甘露肽存在下的表达在72小时后将两种细胞系的细胞活力抑制到40-60%。此外，48 h后HEp-2细胞的膜联蛋白v阳性率和亚g0 /G1期细胞群显著升高（mock vs GALR2分别为12.3 vs 25.0% （P < 0.01）和9.1 vs 32.0% (P < 0.05)）。KB细胞也出现了这些变化，但程度较轻。此外，在HEp-2细胞中，galr2介导的凋亡是caspase不依赖的，包括下调ERK1/2，随后诱导促凋亡的Bcl-2蛋白Bim。

Galanin and its receptors, GALR1 and GALR2, are known tumor suppressors and potential therapeutic targets in head and neck squamous cell carcinoma (HNSCC). 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. Next, we demonstrated that GALR2 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 GALR2: 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, GALR2-mediated apoptosis was caspase-independent, involving downregulation of ERK1/2, followed by induction of the pro-apoptotic Bcl-2 protein, Bim.