Calf Spleen Extractive Injection (CSEI), a small
peptides enriched extraction, performs immunomodulatory activity on
cancer patients suffering from
radiotherapy or
chemotherapy. The present study aims to investigate the anti-
hepatocellular carcinoma effects of CSEI in cells and
tumor-xenografted mouse models. In HepG2 and SMMC-7721 cells, CSEI reduced cell viability, enhanced apoptosis rate, caused
reactive oxygen species (ROS) accumulation, inhibited migration ability, and induced
caspases cascade and mitochondrial membrane potential dissipation. CSEI significantly inhibited HepG2-xenografted
tumor growth in nude mice. In cell and animal experiments, CSEI increased the activations of
pro-apoptotic proteins including
caspase 8,
caspase 9 and
caspase 3; meanwhile, it suppressed the expressions of
anti-apoptotic protein B-cell lymphoma 2 (Bcl-2) and anti-oxidation
proteins, such as nuclear factor-erythroid 2 related factor 2 (Nrf2) and
catalase (CAT). The enhanced phosphorylation of P38 and c-JunN-terminalkinase (JNK), and decreased phosphorylation of extra cellular signal-regulated
protein kinase (ERKs) were observed in CSEI-treated cells and
tumor tissues. CSEI-induced cell viability reduction was significantly attenuated by
N-Acetyl-l-cysteine (a ROS inhibitor) pretreatment. All data demonstrated that the upregulated oxidative stress status and the altered
mitogen-activated protein kinases (MAPKs) phosphorylation contributed to CSEI-driven
mitochondrial dysfunction. Taken together, CSEI exactly induced apoptosis in human
hepatocellular carcinoma cells via ROS/MAPKs dependent mitochondrial pathway.