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Objective:To evaluate the preclinical value of18F-fluoropropionic acid (18F-FPA) and18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) for imaging HCCs.Methods:The18F-FPA and18F-FDG uptake patterns in 3 HCC cell lines (Hep3B, HepG2, and SK-Hep1) were assessed in vitro and in vivo. The18F-FPA uptake mechanism was investigated using inhibition experiments with orlistat and 5-tetradecyloxy-2-furoic acid. The18F-FPA PET imaging was performed in different tumor animal models and compared with18F-FDG. We also evaluated the expressions of glucose transporter-1 (GLUT1), fatty acid synthase (FASN), and matrix metalloproteinase-2 (MMP2) in these cell lines.Results:In vitro experiments showed that the radiotracer uptake patterns were complementary in the HCC cell lines. Orlistat and 5-tetradecyloxy-2-furoic acid decreased the uptake of18F-FPA. The tumor-to-liver ratio of18F-FPA was superior to that of18F-FDG in the SK-Hep1 and HepG2 tumors ( P < .05). However, in the Hep3B tumors, the tumor-to-liver normalized uptake of18F-FDG was higher than18F-FPA ( P < .01). FASN was highly expressed in cell lines with high18F-FPA uptake, whereas GLUT1 was highly expressed in cell lines with high18F-FDG uptake. The18F-FPA uptake correlated with FASN ( r = 0.89, P = .014) and MMP2 ( r = 0.77, P = .002) expressions.Conclusions:PET imaging with18F-FPA combined with18F-FDG can be an alternative for detecting HCC.