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The cytotoxic activity of litchi (Litchi chinensis Sonn.) flower ethanolic extract (LFEE) against A549 (human lung adenocarcinoma), KB (human nasopharyngeal carcinoma), MCF-7 (human breast adenocarcinoma) and HepG2 (human hepatocellular carcinoma) cells was evaluated. LFEE exhibited better anti-proliferative action against HepG2 cells, which induced G2/M phase arrest and apoptosis in HepG2 cells. HepG2 cells treated with LFEE (0.3 mg/mL) enhanced expressions of p53, tBid, Bad and Bax, along with decreased expressions of Bcl-2 and Bcl-xL, and induced the release of cytochrome c from mitochondria to cytosol. That also accompanied by activation of caspases-3, -8 and -9 and cleavage of poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP); furthermore, down-regulations of phosphoinositide 3 kinase (PI3K), Akt, Akt phosphorylation and Bad phosphorylation could also be observed. Five flavonoids (total amount, 101.07 mg/g dried extract (g DE)), nine phenolic acids (total amount, 55.07 mg/ g DE) and proanthocyanidin A2 (81.20 mg/ g DE) could be determined in LFEE.