Prostaglandin E2 promotes the cell growth and invasive ability of hepatocellular carcinoma cells by upregulating c-Myc expression via EP4 receptor and the PKA signaling pathway.

Hepatocellular carcinoma (HCC) represents a major health problem worldwide. Prostaglandin E-2 (PGE(2)), the predominant product of cyclooxygenase-2, has been implicated in hepatocarcinogenesis. However, the underlying molecular mechanisms remain to be further elucidated. c-myc, a cellular proto-oncogene, is activated or overexpressed in many types of human cancer, including HCC. The present study was designed to investigate the internal relationship and molecular mechanisms between PGE(2) and c-Myc in HCC, and to define its role in HCC cell growth and invasion. Our results showed that PGE(2) significantly upregulated c-Myc expression at both the mRNA and protein levels, and knockdown of c-Myc blocked PGE(2)-induced HCC cell growth and invasive ability in human HCC Huh-7 cells. The effect of PGE(2) on c-Myc expression was mainly through the EP4 receptor, and EP4 receptor-mediated c-Myc protein upregulation largely depended on de novo biosynthesis of c-Myc mRNA and its protein. EP4 receptor signaling activated G(s)/AC and increased the intracellular cAMP level in Huh-7 cells. The adenylate cyclase (AC) activator forskolin mimicked the effects of the EP4 receptor agonist on c-Myc expression, while the AC inhibitor SQ22536 reduced EP4 receptor-mediated c-Myc upregulation. These data confirm the involvement of the G(s)/AC/cAMP pathway in EP4 receptor-mediated c-Myc upregulation. Moreover, the phosphorylation levels of CREB protein were markedly elevated by EP4 receptor signaling, and by using specific inhibitor and siRNA interference, we demonstrated that PKA/CREB was also involved in the EP4 receptor-mediated c-Myc upregulation. In summary, the present study revealed that PGE(2) significantly upregulates c-Myc expression at both mRNA and protein levels through the EP4R/G(s)/AC/cAMP/PKA/CREB signaling pathway, thus promoting cell growth and invasion in HCC cells. Targeting of the PGE(2)EP4R/c-Myc pathway may be a new therapeutic strategy to prevent and cure human HCC.