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The marine natural product microsclerodermin A is a novel inhibitor of the nuclear factor kappa B and induces apoptosis in pancreatic cancer cells

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Summary

Pancreatic cancer, the 4th leading cause of cancer death in the US, is highly resistant to all current chemotherapies, and its growth is facilitated by chronic inflammation. An important mediator of inflammation is the nuclear factor kappa B (NFκB), a transcription factor that regulates over 500 genes including the regulation of anti-apoptotic proteins, cell cycle progression and cytokine production. NFκB is constitutively activated in pancreatic cancer cells contributing to their resistance to apoptosis and high metastatic potential. Although many small molecules that inhibit NFκB have been identified, none are currently used in the clinic, perhaps due to their lack of specificity. To identify novel inhibitors of NFκB, the HBOI library of enriched fractions from marine organisms was screened using a reporter cell line that produces luciferin under the transcriptional control of NFκB. Fractions from the sponge Amphibleptula were active in this screen and contained the antifungal cyclic peptide microsclerodermin A. Microsclerodermin A is shown here to inhibit NFκB transcriptional activity in a reporter cell line, to reduce levels of phosphorylated (active) NFκB in the AsPC-1 cell line, to have an IC50 for cytotoxicity in the low micromolar range against the AsPC-1, BxPC-3, MIA PaCa-2 and PANC-1 pancreatic cancer cell lines, and to induce significant apoptosis in the AsPC-1, BxPC-3 and the PANC-1 cell lines. Treatment of AsPC-1 cells with microsclerodermin A also resulted in an increase in IL-8 production without apparent induction of angiogenic factors and there is the possibility that inhibition of NFκB by microsclerodermin A is mediated by the glycogen synthase kinase 3β pathway.

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Grant support

Funding for this project was provided by the State of Florida Biomedical Research Program Bankhead Coley New Investigator Award 09BN-08-23088 (Guzmán) and by NIH R01 CA 093455 and U19 CA 52955 awarded to Dr. Amy Wright. Funding for the student’s stipend (K. Maers) was provided by the Gertrude E. Skelly Charitable Foundation.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Center for Marine Biomedical and Biotechnology Research, Harbor Branch Oceanographic Institute at Florida Atlantic University, 5600 US 1 North, Ft. Pierce, FL, 34946, USA

    Esther A. Guzmán, Kelly Maers, Jill Roberts, Hilaire V. Kemami-Wangun, Dedra Harmody & Amy E. Wright

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  1. Esther A. Guzmán
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  2. Kelly Maers
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  3. Jill Roberts
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  4. Hilaire V. Kemami-Wangun
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  6. Amy E. Wright
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Correspondence to Esther A. Guzmán.

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Notes

This is HBOI contribution number 1941.

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Videos 1

Induction of apoptosis in AsPC-1 cells by Microsclerodermin A. AsPC-1 cells were plated and allowed to adhere overnight in a 96-well clear bottom black plate. Prior to imaging, media was replaced with complete RPMI media with no phenol red that contained 2.4 μM microsclerodermin A (2X IC50 of NFκB inhibition) or its vehicle control methanol, as well as 1 drop NucBlue Live Cell Stain Hoechst 33342 (Molecular Probes R37605) per 1000 mL complete RPMI, 5 μL/well 7-Aminoactinomycin D (7AAD; BD Pharmingen, San Jose California) and 0.75 μL/well NucView 488 Caspase 3 substrate (Biotum Inc., Hayward, CA). Images were captured every 20 min for about 16 h at 20X magnification with an ImageXpress Micro XLS Widefield High-Content Analysis System (Molecular Devices, Sunnyvale, CA). Images were captured in bright field as well to determine cell morphology. The nuclei of all cells were stained with Hoechst. Cells that cleaved caspase 3 show a green color. Those cells that lost membrane integrity allowed the introduction of the 7AAD dye shown in red. Cells undergoing apoptosis should stain green first, followed by red. (AVI 121793 kb)

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Guzmán, E.A., Maers, K., Roberts, J. et al. The marine natural product microsclerodermin A is a novel inhibitor of the nuclear factor kappa B and induces apoptosis in pancreatic cancer cells. Invest New Drugs 33, 86–94 (2015). https://doi.org/10.1007/s10637-014-0185-3

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  • DOI: https://doi.org/10.1007/s10637-014-0185-3

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