Clinical & Experimental Metastasis

, Volume 31, Issue 8, pp 897–907 | Cite as

Capsaicin suppresses the migration of cholangiocarcinoma cells by down-regulating matrix metalloproteinase-9 expression via the AMPK–NF-κB signaling pathway

Research Paper


Cholangiocarcinoma is one of the most difficult malignancies to cure. An important prognostic factor is metastasis, which precludes curative surgical resection. Recent evidence shows that capsaicin has an inhibitory effect on cancer cell migration and invasion. Here, we investigated the molecular mechanism of the capsaicin-induced anti-migration and anti-invasion effects on HuCCT1 cholangiocarcinoma cells. Migration and invasion were significantly reduced in response to capsaicin. Capsaicin also inhibited the expression of matrix metalloproteinase-9 (MMP-9). In capsaicin-treated cells, levels of phosphorylated AMPK increased, and this effect was abolished by treatment with the AMPK inhibitor, Compound C. Capsaicin enhanced the expression of SIRT1, which can activate the transcription factor NF-κB by deacetylation. This suggests that NF-κB is activated by capsaicin via the SIRT1 pathway. In addition, capsaicin-activated AMPK induced the phosphorylation of IκBα and nuclear localization of NF-κB p65. Chromatin immunoprecipitation assays demonstrated that capsaicin reduced MMP-9 transcription by inhibiting NF-κB p65 translocation and deacetylation via SIRT1. These findings provide evidence that capsaicin suppresses the migration and invasion of cholangiocarcinoma cells by inhibiting NF-κB p65 via the AMPK–SIRT1 and the AMPK–IκBα signaling pathways, leading to subsequent suppression of MMP-9 expression.


Capsaicin Migration Cholangiocarcinoma MMP-9 AMPK SIRT1 NF-kB 



Matrix metalloproteinase-9


AMP-activated protein kinase



This work was supported by Priority Research Center Program (2009-0094050) and Basic Research Program (2013R1A1A4A01009559) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology. D-J. Yoon, A-R. Kim is supported by the BK-21 Fund.

Supplementary material

10585_2014_9678_MOESM1_ESM.tif (25 kb)
Supplementary material 1 (TIFF 24 kb) Expression of TIMP-1 and TIMP-2 (endogenous MMP-9 and MMP-2 inhibitors, respectively) is not altered in the capsaicin-treated HuCCT1 cells. Total RNA was extracted from capsaicin-treated cells and RT-PCR was performed using TIMP-1- and TIMP-2-specific primers. Cells were cultured and treated as in Fig. 2
10585_2014_9678_MOESM2_ESM.tif (71 kb)
Supplementary material 2 (TIFF 71 kb) Capsaicin activates SIRT1 by increasing the NAD+/NADH ratio in HuCCT1 cells. Cells were treated with Sirtinol (1 μM) or Compound C (1 μM) followed by 50 μM capsaicin for 24 h. The NAD+/NADH ratio was quantified using an NAD+/NADH Cell-Based Assay Kit (Cayman Chemical Company, MI, USA). The NAD+ and NADH concentrations were determined by colorimetric measurements at 450 nm and NAD+ standard curves. NAD+ concentration was calculated as described in the manufacturer’s manual


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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Gong-Rak Lee
    • 1
  • Soo Hwa Jang
    • 2
  • Chang Jae Kim
    • 2
  • Ah-Ram Kim
    • 2
  • Dong-Joon Yoon
    • 2
  • Neung-Hwa Park
    • 1
    • 2
  • In-Seob Han
    • 3
  1. 1.Department of Medical Science, School of MedicineUniversity of UlsanUlsanKorea
  2. 2.Biomedical Research Center, Ulsan University HospitalUniversity of Ulsan College of MedicineUlsanKorea
  3. 3.Department of Biological SciencesUniversity of UlsanUlsanKorea

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