Abstract
Abstract 4165
Although the exact mechanism of action of thalidomide is still unknown, it has been suggested that it may affect TNF-α in addition to its possible anti-angiogenic and immunomodulatory properties. Studies conducted in our lab have indicated that thalidomide cytotoxicity in KG-1a human acute myelogenous leukemia cell line was enhanced by combining it with arsenic trioxide. So, the current investigation was conducted in order to evaluate the effect of thalidomide either alone or in combination with arsenic trioxide on the release of TNF-α and VEGF from this cell line in an attempt to clarify its possible cytotoxic mechanism. Human acute myelogenous leukemia cell line KG-1a (obtained from American Type Culture Collection, ATCC) grown in complete medium containing Iscove's modified Dulbecco's medium and fetal bovine albumin were used in this study. The cells were cultured for 48 hours in a 12 well-culture plates in duplicates at a concentration of 2×106 cells/ml in the presence or absence of thalidomide (5 mg/L) [Tocris bioscience, Ellisville, Mo] and or arsenic trioxide (4 μM) [Sigma-Aldrich, Inc., St. Louis, MO]. Cells were harvested by centrifugation and the levels of TNF-α and VEGF in the supernatant were determined by ELISA using the Quantikine TNF-α and VEGF kits respectively (R&D Systems, Minneapolis, MN). Results obtained indicate that the levels of TNF-α in the supernatant of KG-1a cell cultures incubated with both thalidomide and arsenic trioxide, whether alone or in combination were statistically lower than those observed in the supernatant of control cells (2.89 pg/ml in thalidomide treated cells supernatant, 5.07 pg/ml in arsenic trioxide treated cells supernatant and 4.15 pg/ml in case of the combined treatment with thalidomide and arsenic trioxide versus 16.88 pg/ml in the supernatant of control cells, p<0.05). However, the levels of VEGF in the supernatant of thalidomide treated cells were statistically higher than those in the supernatant of control cells (69.61 pg/ml versus 11.48 pg/ml, p< 0.001). Of note, that arsenic trioxide whether alone or in combination with thalidomide did not produce any statistically significant difference in the levels of VEGF as compared to control or thalidomide treated cell supernatant. These findings clearly indicate that both thalidomide and arsenic trioxide inhibition of TNF-α production by KG-1a cells may play an important role in their cytotoxic effect. However the increase in VEGF levels in the supernatants of thalidomide treated KG-1a cells may reflect a compensatory mechanism of cells which have survived thalidomide cytotoxicity (Supported by NIH grant RR03020).
Disclosures:
No relevant conflicts of interest to declare.
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