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Characterization of a novel curcumin analog P1 as potent inhibitor of the NF-κB signaling pathway with distinct mechanisms

  
@article{APS7936,
	author = {Yan-min Peng and Jian-bin Zheng and Yu-bo Zhou and Jia Li},
	title = {Characterization of a novel curcumin analog P1 as potent inhibitor of the NF-κB signaling pathway with distinct mechanisms},
	journal = {Acta Pharmacologica Sinica},
	volume = {34},
	number = {7},
	year = {2016},
	keywords = {},
	abstract = {Yan-min PENG1, Jian-bin ZHENG2, Yu-bo ZHOU1, *, Jia LI1, *
1National Center for Drug Screening, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; 2School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China
 
Aim: Curcumin has shown promising anticancer activity, which relies on its inhibition on NF-κB pathway.  In this study, we characterized the pharmacological profile of a novel curcumin analog P1 and elucidate the related mechanisms.
Methods: HEK293/NF-κB cells, stably transfected with an NF-κB-responsive luciferase reporter plasmid, were generated for high-throughput screen (HTS).  Eight cancer cell lines, including PC3, COLO 205, HeLa cells etc.  were tested.  Cell viability was assessed using the sulforhodamine B (SRB) assays.  Cell apoptosis was evaluated using FACS, immunocytochemistry, and Western blotting.  H2-DCFDA and MitoSOX Red were used to detect cellular and mitochondrial reactive oxygen species (ROS).  The mitochondrial function was evaluated using mitochondrial oxygen consumption assay.

Results: P1, a tropinone curcumin, was found in HTS targeting the NF-κB pathway.  Its IC50 value in inhibition of TNF-α-induced NF-κB activation was 0.8 μmol/L, whereas its IC50 values in inhibiting the growth of A549 and HeLa cells were 1.24 and 0.69 μmol/L, respectively, which was 20- to 30-fold more potent than curcumin.  The inhibition of P1 on the NF-κB pathway was further addressed in HeLa cells.  The compound up to 10 µmol/L did not affect the binding of NF-κB to DNA, but markedly inhibited NF-κB nuclear translocation, IκB degradation and IκB kinase phosphorylation.  The compound (1 and 3 µmol/L) concentration-dependently induced ROS generation, whereas curcumin up to 20 µmol/L had no effect.  P1-induced ROS generation was mainly localized in mitochondria, and reversed by NAC.  Moreover, the compound significantly enhanced TNF-α-induced apoptosis. 

Conclusion: P1 is a novel curcumin analog with potent anticancer activities, which exerts a distinct inhibition on the NF-κB pathway.

 
Keywords: curcumin; P1; anticancer agent; high-throughput screen; NF-κB; HeLa cell; mitochondria; ROS
 
This work was supported by grants from the National Natural Science Foundation of China (No 91029716, 81021062, and 81072667).
* To whom correspondence should be addressed.
E-mail jli@mail.shcnc.ac.cn (Jia LI); ybzhou@mail.shcnc.ac.cn (Yu-bo ZHOU)
Received 2012-10-22    Accepted 2013-01-10},
	issn = {1745-7254},	url = {http://www.chinaphar.com/article/view/7936}
}