Eph/Ephrin family, one of the largest receptor tyrosine kinase families, has been extensively studied in morphogenesis and neural development. Recently, growing attention has been paid to its role in the initiation and progression of various cancers. However, the role of Eph/Ephrins in hepatocellular carcinoma (HCC) has been rarely investigated. In this study, we found that the expression of EphrinA2 was significantly up-regulated in both established cell lines and clinical tissue samples of HCC, and the most significant increase was observed in the tumors invading the portal veins. Forced expression of EphrinA2 in HCC cells significantly promoted in vivo tumorigenicity, whereas knockdown of this gene inhibited this oncogenic effect. We further found that suppression of apoptosis, rather than accelerating proliferation, was responsible for EphrinA2-enhanced tumorigenicity. In addition, EphrinA2 endowed cancer cells with resistance to tumor necrosis factor alpha (TNF-␣)-induced apoptosis, thus facilitating their survival. Furthermore, we disclosed a novel EphrinA2/ras-related c3 botulinum toxin substrate 1 (Rac1)/V-akt murine thymoma viral oncogene homolog ( H epatocellular carcinoma (HCC) is a major health problem worldwide, ranking as the fifth most common cancer in the world and the third most common cause of cancer-related death. 1 HCC shows great geographical variation, with a very high incidence in Asia and sub-Saharan Africa, but it is now becoming more common in the West. During the past 20 years in the United States, HCC has risen by 114%. 2 This paralleled an increase in the incidence of chronic hepatitis, which serves as a main risk factor for HCC. 3 At the initiation of hepatic oncogenesis, transformed hepatocytes must elude various cellular defense activities and acquire abnormal capabilities to survive and proliferate. 4 Aberrant signaling through receptor tyrosine kinases plays a pivotal role in the development and progression of HCC. 5 Eph receptors constitute one of the largest receptor tyrosine kinase families and have been reported to be involved in a variety of cancers. For example, up-regulation of EphA2 has been observed in many malignant tumors [6][7][8] and is associated with accelerated cell prolifera-