Acta Pharmacologica Sinica (2009) 30: 134-140; doi: 10.1038/aps.2008.13; published online 22nd December 2008

In vitro anticancer property of a novel thalidomide analogue through inhibition of NF-κB activation in HL-60 cell

Aim: To investigate the anticancer property and possible mechanism of action of a novel sugar-substituted thalidomide derivative (STA-35) on HL-60 cells in vitro. 

Methods: TNF-α-induced NF-κB activation was determined using a reporter gene assay.  The MTT assay was used to measure cytotoxicity of the compound.  The appearance of apoptotic Sub-G1 cells was detected by flow cytometry analysis.  PARP cleavage and protein expression of NF-κB p65 and its inhibitor IκB were viewed by Western blotting.

Results: STA-35 (1?0 µmol/L) suppressed TNF-α-induced NF-κB activation in transfected cells (HEK293/pNiFty-SEAP) in a dose- (1?0 µmol/L) and time-dependent (0?8 h) manner.  It was also shown that STA-35 exerted a dose-dependent inhibitory effect on HL-60 cell proliferation with an IC50 value of 9.05 μmol/L.  In addition, STA-35 induced apoptosis in HL-60 cells, as indicated by the appearance of a Sub-G1 peak in the cell cycle distribution, as well as poly ADP-ribose polymerase (PARP) cleavage.  Subsequently, both NF-κB p65 and its inhibitor IκB gradually accumulated in cytoplasmic extracts in a dose- and time-dependent manner, indicating the blockage of NF-κB translocation induced by TNF-α from the cytoplasm to the nucleus. 

Keywords: anticancer; HL-60; thalidomide; NF-κB; apoptosis