Medications targeting the somatostatin type 2 receptor (SSTR2) have been employed for pancreatic inflammations and cancers, possibly via the regulation of the transcription factor nuclear factor B (NFB). Here we demonstrate that in pancreatic acinar AR42J cells, activation of SSTR2 leads to the stimulation of inhibitor B kinase (IKK)/NFB signaling cascade via PTX-insensitive G proteins in a time- and dose-dependent manner. The inability of G_q/11 and G_12/13 proteins to activate IKK/NFB by SSTR2 in transfected human embryonic kidney (HEK) 293 cells and the lack of G₁₆ in AR42J cells suggested a possible role of G₁₄ in mediating the SSTR2-induced responses. This regulatory role of G₁₄ was further confirmed by the activation of IKK and NFB in HEK 293 cells expressing SSTR2 and G₁₄ upon induction. The stimulatory effect of G12 and the abrogation by overexpressing transducin confirmed the participation of G in SSTR2-mediated IKK/NFB activation. By the application of specific inhibitors and dominant negative mutants, phospholipase C, protein kinase C and calmodulin-dependent kinase II were shown to be involved in the SSTR2-induced responses. Inhibition of c-Src and numerous intermediates, including Ras, Raf-1 kinase, MEK1/2, along the extracellular signal-regulated kinase (ERK) cascade attenuated the somatostatin-mediated IKK/NFB activation. Although c-Jun N-terminal kinase and p38 MAPK were also stimulated by SSTR2, suppression of these two MAPKs was ineffective in altering the somatostatin-mediated responses. Similar results were also obtained using AR42J cells. These data suggest that the activation of IKK/NFB signaling cascade by SSTR2 requires a complicated network consisting of G(sub14} and multiple intermediates.