Cytobine-induced killer cells promote apoptosis of human liver cancer stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Immunotherapy with autologous immune cel s has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated. OBJECTIVE:To investigate the relationship between cytokine-induced kil er cel secretion and apoptosis in human liver cancer stem cel s. METHODS:Human liver cancer stem cel s, HepG2 cel s, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cel s from patients with liver cancer were induced byγ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form kil er cel s. Passage 1 liver cancer stem cel s were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced kil er cel s and human liver cancer stem cel s). At 48 hours after culture, apoptosis in human liver cancer stem cel s was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively. RESULTS AND CONCLUSION:The apoptotic rate in the control group was significantly lower than that in the experimental group (P<0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P<0.05). Experimental findings show that cytokines-induced kil er cel s can significantly promote apoptosis in human liver cancer stem cel s, and up-regulate the caspase-3 mRNA and protein expressions dramatical y.