沙利度胺联合干扰素对Kasumi-1细胞的增殖抑制作用及其机制研究

OBJECTIVE: To explore the inhibitory effect of thalidomide combined with interferon (IFN) on the human acute myeloid leukemia cell line Kasumi-1 and its mechanism. METHODS: The inhibitiory effect of Kasumi-1 cells by thalidomide, interferon or combination was detected by CCK-8 method, the apoptosis by flow cytometry, the expression of apoptosis related proteins by Western blot, vascular endothelial growth factor (VEGF) concentration in culture supernatant by ELISA. RESULTS: Thalidomide inhibited the proliferation of Kasumi-1 in a dose-dependent manner from 50 µg/ml to 500 µg/ml with an IC(50) of (451.13±6.92) µg/ml at 24 h and (362.50±14.52) µg/ml at 48 h. IFN also demonstrated the inhibitory capacity in a dose-dependent manner from 500 U/ml to 5 000 U/ml, with an IC(50) of (2 209±127) U/ml at 24 h and (1 393±63) U/ml at 48 h. The apoptosis rates of Kasumi-1 cells treated with thalidomide 350 µg/ml or IFN 1 400 U/ml for 48 h were (14.68±2.61)% and (21.71±0.71)%, respectively, significantly higher than control group (P<0.01). In combination group the inhibition and the apoptosis rate were (88.50±2.40)% and (41.95±3.41)%, significantly higher than control and each single agent group (P<0.01). The VEGF concentrations of combination group [(94.61±5.46) ng/L] decreased significantly, as compared to thalidomide group [(141.11±3.70) ng/L] and IFN group [(119.90±2.00) ng/L] (P<0.05). Western blot analysis showed Bcl-2 expression of Kasumi-1 cells decreased, while p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 increased after treated with thalidomide 350 µg/ml or IFN 1 400 U/ml for 48 h. When treated with the combination agents, the expression of Bcl-2 further decreased and p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 further increased as compared with each single agent (P<0.05). CONCLUSION: Thalidomide and IFN could synergistically inhibit the proliferation of Kasumi-1 cells probably through inducing apoptosis via the mitochondrial pathway, death receptor pathway and P38 MAPK pathway, as well as inhibiting VEGF secretion.