Growth-related oncogene (GRO), a member of the CXC chemokine subfamily, plays a major role in inflammation and wound healing. CXC chemokines have been found to be associated with tumorigenesis, angiogenesis, and metastasis. Although elevated expression of GRO has been reported in several human cancers, the expression and role of GRO and its receptor, CXCR2, in esophageal cancer are poorly understood. This study used real-time reverse transcription-PCR (RT-PCR) and immunohistochemical approaches to show that GROα, GROβ, and CXCR2 are up-regulated in esophageal tumor tissue. Furthermore, GROα, GROβ, and CXCR2 are constitutively expressed in WHCO1, an esophageal cancer cell line that was used as a model system here. GROβ enhances transcription of EGR-1, via the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway, which can be blocked by a specific antagonist of CXCR2 (SB 225002) or specific antibody to GROβ. WHCO1 cells treated with SB 225002 exhibited a 40% reduction in cell proliferation. A stable WHCO1 GROα RNA interference (RNAi) clone displayed a 43% reduction in GROα mRNA levels as determined by real-time RT-PCR, reduced levels of GROα by fluorescence microscopy, and a 60% reduction in the levels of phosphorylated ERK1/2. A stable clone expressing GROβ RNAi displayed >95% reduction in GROβ mRNA levels, reduced levels of GROβ by fluorescence microscopy, and an 80% reduction in the levels of phosphorylated ERK1/2. Moreover, these GROα RNAi- and GROβ RNAi-expressing clones displayed a 20% and 50% decrease in cell proliferation, respectively. Our results suggest that GROα-CXCR2 and GROβ-CXCR2 signaling contributes significantly to esophageal cancer cell proliferation and that this autocrine signaling pathway may be involved in esophageal tumorigenesis. (Cancer Res 2006; 66(6): 3071-7)