Water-soluble 4-(dimethylaminomethyl)heliomycin exerts greater antitumor effects than parental heliomycin by targeting the tNOX-SIRT1 axis and apoptosis in oral cancer cells
- Institute of Biomedical Sciences, National Chung Hsing University, Taiwan
- Institute of Medicine, Chung Shan Medical University, Taiwan
- Department of Otorhinolaryngology-Head and Neck Surgery, Changhua Christian Hospital, Taiwan
- Department of Post-Baccalaureate Medicine, College of Medicine, National Chung Hsing University, Taiwan
- Gause Institute of New Antibiotics, Russian Federation
- Department of Medical Research, China Medical University Hospital, Taiwan
- Graduate Institute of Basic Medicine, China Medical University, Taiwan
Figures
Figure 1
Structures of heliomycin and 4-(dimethylaminomethyl)heliomycin (designated 4-dmH).
Figure 2
CETSA-based determination of binding between 4-dmH and SIRT1 protein.
(a) Cells were incubated with different concentrations of 4-dmH as described in the Methods. Dose-dependent thermal stabilization of SIRT1 was assessed after heating samples at 54 °C for 3 min in …
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Figure 2—source data 1
Original files for the western blot analysis in Figure 2.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig2-data1-v1.zip
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Figure 2—source data 2
Tiff files containing Figure 2 of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig2-data2-v1.zip
Figure 3
CETSA-based determination of binding between heliomycin and SIRT1 protein.
(a) Cells were incubated with different concentrations of heliomycin as described in the Methods. Dose-dependent thermal stabilization of SIRT1 was assessed after heating samples at 54 °C for 3 min …
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Figure 3—source data 1
Original files for the western blot analysis in Figure 3.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig3-data1-v1.zip
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Figure 3—source data 2
Tiff files containing Figure 3 of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig3-data2-v1.zip
Figure 4
SIRT1 inhibition and downregulation by water-soluble 4-dmH.
(a) SIRT1 activity was determined by a SIRT1 Activity Assay Kit (Fluorometric) with control or recombinant protein treated with test compounds. Values (mean ±SE) are from three independent …
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Figure 4—source data 1
Original files for the western blot analysis in Figure 4.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig4-data1-v1.zip
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Figure 4—source data 2
PDF containing Figure 4b-d of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig4-data2-v1.zip
Figure 5
Heliomycin and 4-dmH provoked different cell death pathways in oral cancer cells.
(a) Cells were exposed to heliomycin or 4-dmH and the percentage of the autophagic population was determined by AO staining using flow cytometry. The results are expressed as a percentage of …
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Figure 5—source data 1
The flow cytometry dot blot autophagy data in Figure 5a.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig5-data1-v1.zip
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Figure 5—source data 2
The flow cytometry dot blot JC-1 staining data in Figure 5b.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig5-data2-v1.zip
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Figure 5—source data 3
Original files for the western blot analysis in Figure 5.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig5-data3-v1.zip
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Figure 5—source data 4
PDF containing Figure 5c,d of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig5-data4-v1.zip
Figure 6
The effects of heliomycin and 4-dmH on proliferation, colony formation, and cell cycle progression of oral cancer cells.
(a, b) Cell proliferation was dynamically monitored by impedance measurements in SAS, HSC-3 cells, and BSEA-2B cells as described in the Methods. Shown are the normalized cell index values measured. …
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Figure 6—source data 1
The colony formation representative data in Figure 6c.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig6-data1-v1.zip
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Figure 6—source data 2
The flow cytometry dot blot cell cycle analysis data in Figure 6d.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig6-data2-v1.zip
Figure 7
tNOX inhibition and downregulation by water-soluble 4-dmH.
(a) The intracellular NAD+/NADH ratio was measured by an NADH/NAD Quantification Kit (BioVision Inc) with control or lysates of SAS cells treated with heliomycin or 4-dmH. Values (mean ±SE) are from …
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Figure 7—source data 1
Original files for the western blot analysis in Figure 7.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig7-data1-v1.zip
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Figure 7—source data 2
PDF containing Figure 7b-e of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig7-data2-v1.zip
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Figure 7—source data 3
The flow cytometry dot blot autophagy and apoptosis data in Figure 7e.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig7-data3-v1.zip
Figure 8
CETSA-based determination of binding between 4-dmH and tNOX protein.
(a) Cells were incubated with different concentrations of the 4-dmH as described in the Methods. Dose-dependent thermal stabilization of tNOX was assessed after heating samples at 54 °C for 3 min in …
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Figure 8—source data 1
Original file for the western blot analysis in Figure 8.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig8-data1-v1.zip
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Figure 8—source data 2
PDF containing Figure 8 of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig8-data2-v1.zip
Figure 9 with 1 supplement
The binding modes of heliomycin and 4-dmH after docking into the pocket of SIRT1 (a, b) and tNOX (c, d).
(a) Superimposition of the docked heliomycin (red) and 4-dmH (blue). (b) Schematic presentations of possible interactions between test compounds and SIRT1 residues. (c) Superimposition of the docked …
Figure 9—figure supplement 1
The seven interaction residues on tNOX were substituted with alanine or glycine amino acids and then simulated the protein structures.
The simulated tNOX structures (a, b) and the binding modes of 4-dmH after docking study (c, d). (a) Superimposition of three types of tNOX structures, including the original tNOX structure (orange) …
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Figure 9—figure supplement 1—source data 1
The simulated tNOX structures (a, b) and the binding modes of 4-dmH after docking study (c, d).
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig9-figsupp1-data1-v1.zip
Figure 10
Therapeutic potential of heliomycin and 4-dmH in oral cancer.
(a–c) In a tumor-bearing mouse xenograft model, control mice were intratumorally injected with vehicle buffer and treatment group mice were intratumorally treated with heliomycin or 4-dmH as …
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Figure 10—source data 1
Original files for the western blot analysis in Figure 10.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig10-data1-v1.zip
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Figure 10—source data 2
PDF containing Figure 10c, d of the relevant western blot analysis with the uncropped gels or blots with the relevant bands.
- https://cdn.elifesciences.org/articles/87873/elife-87873-fig10-data2-v1.zip
Author response image 1
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The simulated tNOX structures (a, b) and the binding modes of 4-dmH after docking study (c, d).
(a) Superimposition of three types of tNOX structures, including the original tNOX structure (orange) and the critical residues in tNOX protein substituted with alanine (magenta) or glycine (cyan). …
Author response image 6
