Abstract
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 (1–20 μmol/L) suppressed TNF-α-induced NF-κB activation in transfected cells (HEK293/pNiFty-SEAP) in a dose- (1–20 μmol/L) and time-dependent (0–48 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.
Conclusion:
A novel sugar-substituted thalidomide derivative, STA-35, is potent toward HL-60 cells in vitro and induces apoptosis by the suppression of NF-κB activation.
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Abbreviations
- DMSO:
-
dimethyl sulfoxide
- NF-κB:
-
nuclear factor-κB
- IκB:
-
Inhibitor of κB
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- TNF-α:
-
tumor necrosis factor-α
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Acknowledgements
The present work was supported by grants from the National High Technology Development Project (863 project, No 2004AA2Z3783) and the National Natural Sciences Foundation of China (No 30330690, 30672525).
We are grateful to Dr Davriche (INSERM 563, CHU Purpan, Toulouse, France) for providing the transfected cell line, HEK293/pNiFty-SEAP.
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Li, M., Sun, W., Yang, Yp. et al. In vitro anticancer property of a novel thalidomide analogue through inhibition of NF-κB activation in HL-60 cells. Acta Pharmacol Sin 30, 134–140 (2009). https://doi.org/10.1038/aps.2008.13
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DOI: https://doi.org/10.1038/aps.2008.13
