Journal: Translational Cancer Research

Article Title: Albiflorin enhances the antitumor effects of propofol in hepatocellular carcinoma cells by inhibiting the Wnt/β-catenin signaling activation

doi: 10.21037/tcr-2025-1246

Figure Lengend Snippet: Combined effects of albiflorin and propofol on the proliferation and apoptosis of Huh-7 cells. (A) Cell viability detected by MTT assay after treatment with propofol alone or in combination with albiflorin for 48 h; (B,C) apoptosis rate assessed by flow cytometry; (D-F) protein expression levels of Bax and Bcl-2 determined by western blot. * P<0.05, *** P<0.001, **** P<0.0001 vs. control group; ## P<0.01, #### P<0.0001 vs. 120 μM propofol treatment group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

Article Snippet: Huh-7 cells were seeded into appropriate culture plates and treated with different concentrations of albiflorin (0, 25, 50, 100 μg/mL; HY-N0037, Purity: 98.67%, MedChemExpress) or propofol (0, 30, 60, 120 μM) individually, or with a combination of 100 μg/mL albiflorin and 120 μM propofol.

Techniques: MTT Assay, Flow Cytometry, Expressing, Western Blot, Control

Journal: Translational Cancer Research

Article Title: Albiflorin enhances the antitumor effects of propofol in hepatocellular carcinoma cells by inhibiting the Wnt/β-catenin signaling activation

doi: 10.21037/tcr-2025-1246

Figure Lengend Snippet: Combined effects of albiflorin and propofol on the migration, invasion, and EMT of Huh-7 cells. Huh-7 cells were treated with propofol alone or in combination with albiflorin for 48 h. (A,B) Cell migration ability was evaluated by Transwell assay, and cells were stained with 0.1% crystal violet; (A,C) cell invasion ability was evaluated by Transwell assay, and cells were stained with 0.1% crystal violet; (D-F) western blot analysis of N-cadherin and E-cadherin in Huh-7 cells. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 vs. control group; # P<0.05, ## P<0.01, #### P<0.0001 vs. 120 μM propofol treatment group. EMT, epithelial-mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Huh-7 cells were seeded into appropriate culture plates and treated with different concentrations of albiflorin (0, 25, 50, 100 μg/mL; HY-N0037, Purity: 98.67%, MedChemExpress) or propofol (0, 30, 60, 120 μM) individually, or with a combination of 100 μg/mL albiflorin and 120 μM propofol.

Techniques: Migration, Transwell Assay, Staining, Western Blot, Control

Journal: Translational Cancer Research

Article Title: Albiflorin enhances the antitumor effects of propofol in hepatocellular carcinoma cells by inhibiting the Wnt/β-catenin signaling activation

doi: 10.21037/tcr-2025-1246

Figure Lengend Snippet: Combined effects of albiflorin and propofol on the Wnt/β-catenin signaling pathway of Huh-7 cells. Huh-7 cells were treated with propofol alone or in combination with albiflorin for 48 h. (A-C) The protein levels of Wnt3a and β-catenin in Huh-7 cells treated with different concentrations of albiflorin were examined using western blotting; (D,E) the mRNA levels of β-catenin and Wnt3a in Huh-7 cells treated with different concentrations of albiflorin were examined using RT-qPCR; (F-H) the protein levels of cyclin D1 and c-Myc in Huh-7 cells were examined using western blotting; (I,J) the mRNA levels of cyclin D1 and c-Myc in Huh-7 cells were examined using RT-qPCR. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 vs. control group; # P<0.05, ## P<0.01, ### P<0.001, #### P<0.0001 vs. 120 μM propofol treatment group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; mRNA, messenger RNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Huh-7 cells were seeded into appropriate culture plates and treated with different concentrations of albiflorin (0, 25, 50, 100 μg/mL; HY-N0037, Purity: 98.67%, MedChemExpress) or propofol (0, 30, 60, 120 μM) individually, or with a combination of 100 μg/mL albiflorin and 120 μM propofol.

Techniques: Western Blot, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Biochemistry

Article Title: Substrate-Dependent Modulation of SIRT2 by a Fluorescent Probe, 1-Aminoanthracene.

doi: 10.1021/acs.biochem.0c00564

Figure Lengend Snippet: Figure 1. SIRT2 interacts with AMA and propofol at the same site. (A) Chemical structures of AMA and propofol. (B) Emission spectra of 100 nM AMA equilibrated with the indicated SIRT2 concentrations in PBS. The excitation wavelength was 380 nm. (C) Fluorescence emission intensities at 520 nm were taken from the data in panel B and plotted vs the SIRT2 concentration to derive a binding isotherm. (D) Emission spectra of 100 nM AMA equilibrated with 4 μM SIRT2 and varying propofol concentrations. Propofol caused a dose-dependent decrease in AMA fluorescence and a red-shift of the spectrum, which indicates AMA unbinding SIRT2 and returning to the aqueous solution. (E) Emission intensities at 520 nm were taken from the data in panel D to derive a dose−response curve for propofol’s reduction of AMA’s fluorescence. The IC50 of 27 μM corresponds to a Kd of 25 μM for SIRT2’s interaction with propofol.29 (F) Interaction of SIRT2(Y139G) with AMA was measured by changes in AMA fluorescence at multiple protein concentrations, as in panel C. In panel F, the Kd is approximate because all measurements were made with protein concentrations below the Kd, which is prior to significant curve saturation.

Article Snippet: AMA (1-aminoanthracene, >98% purity) was purchased from TCI, and propofol (2,6-diisopropylphenol, 97% purity) was purchased from Alfa Aesar.

Techniques: Fluorescence, Concentration Assay, Binding Assay

Journal: Biochemistry

Article Title: Substrate-Dependent Modulation of SIRT2 by a Fluorescent Probe, 1-Aminoanthracene.

doi: 10.1021/acs.biochem.0c00564

Figure Lengend Snippet: Figure 2. Varying effects of acylated peptides on interactions of SIRT2 with AMA. (A) Chemical structures of lysine acyl modifications on H4K16 peptides. (B) Myristoyl-H4K16 peptide inhibited binding of AMA to SIRT2, which was measured by the decreased AMA fluorescence in the presence of the peptide. The IC50 of 3 μM corresponds to a Kd of 2.7 μM for SIRT2’s interaction with myristoyl-H4K16 peptide. (C) Inhibition of AMA binding to SIRT2 caused by dodecanoyl-H4K16 peptide. (D) Decanoyl-H4K16 peptide significantly enhanced the binding of AMA to SIRT2. Note the different y-axis scale on this and other panels. (E) Enhancement of AMA binding to SIRT2 mediated by 4-oxononanoyl-H4K16 peptide. (F) AMA’s affinity for SIRT2 in the presence of 10 μM decanoyl-H4K16 peptide was measured by its change in fluorescence with increasing SIRT2 concentrations. (G) Propofol inhibited binding of AMA to SIRT2 in the presence of 10 μM decanoyl-H4K16 peptide. The IC50 of 8.6 μM corresponds to a Kd of 7 μM for propofol’s interaction with SIRT2 in the presence of 10 μM decanoyl-H4K16 peptide. (H) Decanoyl- H4K16 peptide enhanced the binding of AMA to SIRT2(Y139G), but to a lesser extent than wild-type SIRT2 (see panel D). (I) Acetyl-H4K16 peptide had no effect on the binding of AMA to SIRT2, and thus no change in fluorescence was observed. (J) Slight increase in the level of AMA binding to 4 μM SIRT2 when the protein was equilibrated with 100 μM acetyl-H4K16 peptide, 300 μM nicotinamide, and 500 μM ADP-ribose. (K) Weak binding of AMA to 10 μM SIRT6 alone, and no change in binding when 300 μM ADP-ribose or 100 μM decanoyl-H4K16 peptide was added. For all panels, the AMA concentration was 100 nM.

Article Snippet: AMA (1-aminoanthracene, >98% purity) was purchased from TCI, and propofol (2,6-diisopropylphenol, 97% purity) was purchased from Alfa Aesar.

Techniques: Binding Assay, Inhibition, Concentration Assay

Journal: Biochemistry

Article Title: Substrate-Dependent Modulation of SIRT2 by a Fluorescent Probe, 1-Aminoanthracene.

doi: 10.1021/acs.biochem.0c00564

Figure Lengend Snippet: Figure 3. Peptide-induced structural changes in the AMA binding site. (A) Co-crystal structure of SIRT2 and myristoyl-peptide [Protein Data Bank (PDB) entry 4Y6L]. The myristoyl modification (green sticks) projects into the AMA/propofol binding site. Tyr139, Phe190, and Leu206 are shown as red spheres. (B) Decanoyl-peptide bound to SIRT2, modeled from PDB entry 4Y6L. The decanoyl modification (green sticks) does not quite reach the back of the hydrophobic tunnel, allowing the formation of a 182 Å3 pocket at the AMA/propofol binding site. The pocket is colored blue. (C) Dodecanoyl-peptide bound to SIRT2, modeled as in panel B, with a 102 Å3 pocket shown at the end of the tunnel. (D) Co- crystal structure of SIRT2, 4-oxononanoyl-peptide, and carba-NAD+ (PDB entry 5G4C). The volume of the pocket at the AMA/propofol binding site is 224 Å3 in this structure, and for perspective, the pocket is 6 Å from the nearest carba-NAD+ atom. Carba-NAD+ would be positioned identically in all of the peptide-bound structures. (E) Co-crystal structure of SIRT2 and trifluoroacetyl-peptide (PDB entry 5FYQ). Select residues that form SIRT2’s hydrophobic tunnel are shown as sticks. Colored red are Tyr139, Phe190, and Leu206, and the electron density of these residues is shown as blue mesh. Note how Tyr139 is not packed with the other residues. In this panel and panels F and G, the contour level of the electron density maps is 0.5σ. (F) Co-crystal structure of SIRT2 and 4-oxononanoyl-peptide (PDB entry 5G4C), with carba-NAD+ removed for the sake of clarity. Note the stronger electron density surrounding Tyr139 compared to that in panel E and its repositioning near the end of the hydrophobic tunnel. (G) Co-crystal structure of SIRT2 and myristoyl-peptide (PDB entry 4Y6L). Residues Tyr139, Phe190, and Leu206 fully form the back of the tunnel and are stabilized (i.e., have stronger electron density) compared to structures in panels E and F.

Article Snippet: AMA (1-aminoanthracene, >98% purity) was purchased from TCI, and propofol (2,6-diisopropylphenol, 97% purity) was purchased from Alfa Aesar.

Techniques: Binding Assay

Journal: Biochemistry

Article Title: Substrate-Dependent Modulation of SIRT2 by a Fluorescent Probe, 1-Aminoanthracene.

doi: 10.1021/acs.biochem.0c00564

Figure Lengend Snippet: Figure 4. AMA and propofol differentially modulate SIRT2’s deacylase activities. (A) Kinetics of SIRT2 deacetylase activity using the tryptophan- labeled acetyl-H4K16 peptide as the substrate. The buffer contained PBS with 1 mM NAD+ and 1 mM DTT, and reactions were run at 37 °C. (B) Kinetics of SIRT2 deacylation of the tryptophan-labeled 4-oxononanoyl-H4K16 peptide under conditions identical to those of panel A. (C) Kinetics of SIRT2 dedecanoylase activity using the tryptophan-labeled decanoyl-H4K16 peptide. (D) Kinetics of SIRT2 demyristoylase activity using the tryptophan-labeled myristoyl-H4K16 peptide. (E) Inhibition of SIRT2 deacetylase activity by AMA and propofol using the indicated peptide substrate and conditions identical to those of panel A. (F) Enhancement and inhibition of SIRT2 deacylase activity by AMA and propofol, respectively, using the H4K16 peptide substrate containing a 4-oxononanoyl modification. (G) Inhibition and enhancement of SIRT2 dedecanoylase activity by AMA and propofol, respectively. Dedecanoylase assays were run using a substrate concentration that approximated the Km (left), and also under kcat conditions (right bar graph). (H) Inhibition of SIRT2 demyristoylase activity by AMA and propofol.

Article Snippet: AMA (1-aminoanthracene, >98% purity) was purchased from TCI, and propofol (2,6-diisopropylphenol, 97% purity) was purchased from Alfa Aesar.

Techniques: Histone Deacetylase Assay, Activity Assay, Labeling, Inhibition, Concentration Assay

Journal: iScience

Article Title: Direct visualization of general anesthetic propofol on neurons by stimulated Raman scattering microscopy

doi: 10.1016/j.isci.2022.103936

Figure Lengend Snippet:

Article Snippet: Propofol-d17 , Toronto Research Chemicals , P829752.

Techniques: Recombinant, Software