Acta Pharmacologica Sinica (2010) 31: 81–92; doi: 10.1038/aps.2009.182

Jin-yi WU^1,#, Kun-wei TSAI^2,#, Jia-jen SHEE³, Yi-zhen LI¹, Ching-hsein CHEN¹, Jing-jing CHUANG⁴, Yi-wen LIU^1,* 

¹Graduate Institute of Biomedical and Biopharmaceutical Sciences, ⁴Department of Microbiology and Immunology, College of Life Sciences, National Chiayi University, Chiayi, Taiwan 60004, China; ²Department of Internal Medicine, Buddhist Tzuchi Dalin General Hospital, Dalin Town, Chiayi, Taiwan 62247, China; ³Division of Urology, Department of Surgery, Chang Gung Memorial Hospital, Chiayi, Taiwan 61363, China

Aim: To examine the antitumor effect of 4′-chloro-3,5-dihydroxystilbene, a resveratrol derivative, on lung adenocarcinoma A549 cells.  

Methods: The cytotoxic IC₅₀ was determined by direct cell counting.  Flow cytometry, monodansylcadaverine (MDC) staining, transfection, Western blot and a proteasome activity assay were used to study the cellular mechanism of 4′-chloro-3,5-dihydroxystilbene.  A xenograft nude mouse model was used to analyze the antitumor effect in vivo.

Results: 4′-Chloro-3,5-dihydroxystilbene induced a rapid and persistent increase in the intracellular reactive oxygen species in the cells, but the cell death could not be inhibited by two antioxidant agents.  The derivative caused sub-G₁ formation, a decrease in the mitochondria membrane potential and poly (ADP-ribose) polymerase degradation, and the caspase inhibitor Z-VAD-FMK could partially prevent cell death.  It also induced a significant increase in intracellular acidic vacuoles, LC3-II formation and intracellular GFP-LC3 aggregation. An autophagic inhibitor partially reversed cell death.  Additionally, 4′-chloro-3,5-dihydroxystilbene induced the accumulation of ubiquitinated conjugates and inhibited proteasome activity in cells.  In an in vivo study, 4′-chloro-3,5-dihydroxystilbene retarded tumor growth in nude mice.

Conclusion: These data suggest that the resveratrol derivative 4′-chloro-3,5-dihydroxystilbene could be developed as an anti-tumor compound.

Keywords: 4′-chloro-3,5-dihydroxystilbene; resveratrol derivative; antitumor effects; apoptosis; lung adenocarcinoma A549 cells