Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Cytotoxic effects of 12 benzocycloheptene analogous drugs toward human hepatoma cells.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques:

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Dose- and time-dependent effects of deptropine on the cell viability of human hepatoma cells. ( a ) Hep3B and ( b ) HepG2 cells were treated with different concentrations of deptropine for 24, 48, or 72 h, and cell viability was determined by an 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Data are presented as the mean ± standard error of three independent experiments.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: MTT Assay

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Effects of deptropine on marker protein expressions of endoplasmic reticular stress and autophagy, as well as the formation of light chain 3 (LC3) puncta, in human hepatoma cells. ( a , b ) Hep3B and HepG2 cells were treated with different concentrations of deptropine for 48 h, and the marker protein expressions of ( a ) endoplasmic reticular stress and ( b ) autophagy were determined by Western blotting. The relative intensity of each band (indicated below the bands) was normalized to the total protein (including protein kinase RNA-like endoplasmic reticulum kinase (PERK), eukaryotic translation initiation factor 2 subunit alpha (eIF2α), and protein kinase B (PKB, also known as Akt) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) loading control. Grp78, glucose-regulated protein 78; AMPK, adenosine 5′-monophosphate-activated protein kinase; SQSTM1, sequestosome-1; ATG7, autophagy related 7; VPS34, vacuolar protein sorting 34. ( c ) Hep3B and HepG2 cells were treated with different concentrations of deptropine for 6 or 12 h, and light chain 3B (LC3B) expression was determined by Western blotting. The relative intensity of LC3B-II was normalized to the GAPDH loading control and indicated below the bands. ( d , e ) HepG2 and Hep3B cells were treated with 20 μM of deptropine for 24 h, and LC3 puncta were detected by immunofluorescence staining with an LC3-specific antibody (green) and nucleic acid staining with 4′,6-diamidino-2-phenylindole (DAPI) (blue). ( d ) Representative immunofluorescence images are shown, and ( e ) quantification of LC3 puncta per cell is presented as the mean ± standard error. * p < 0.01.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: Marker, Western Blot, Control, Expressing, Immunofluorescence, Staining

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Effects of deptropine on inhibition of autophagosome-lysosome fusion in human hepatoma cells. Hep3B cells were transfected with the FUW mCherry-green fluorescent protein (GFP)-LC3 plasmid and then treated with 20 μM of deptropine for 24 h. Autophagosomes/autolysosomes were visualized with fluorescence microscopy. Chloroquine (25 μΜ) was used as a positive control. ( a ) Representative fluorescence images are shown, and ( b ) autophagic flux was estimated as the ratio between yellow-positive cells and red-positive cells, which are presented as the mean ± standard error. * p < 0.0001.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: Inhibition, Transfection, Plasmid Preparation, Fluorescence, Microscopy, Positive Control

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Effects of deptropine on the lysosomal activity of human hepatoma cells. ( a ) Hep3B and ( b ) HepG2 cells were treated with different concentrations of deptropine for 24 and 48 h. Total cell lysates were collected, and the precursor form (pre-), intermediate (i-), and mature (m-) form of the cathepsin L (CTSL) protein were detected by Western blotting. The relative intensity of pre-CTSL was normalized to the GAPDH loading control and indicated below the bands.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: Activity Assay, Western Blot, Control

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Effects of deptropine on caspase expressions and activities in human hepatoma cells. Hep3B and HepG2 cells were treated with different concentrations of deptropine for 48 h, and ( a ) poly(ADP ribose) polymerase (PARP), caspase-3, -8, and -9 protein expressions were determined by Western blotting. The relative intensity of each band (indicated below the bands) was normalized to the GAPDH loading control. ( b ) The cell extracts were subjected to caspase-3, -8, and -9 activity assays as described in “Materials and Methods”. Values were obtained in two independent experiments performed in triplicate, and results are presented as the mean ± standard error. * p < 0.05 vs. the control. STS, staurosporine (1 μM) as the positive control.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: Western Blot, Control, Activity Assay, Positive Control

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: Effects of deptropine on hepatoma tumor xenografts in nude mice. Hep3B cells were subcutaneously injected between the scapulas of athymic nude mice, and the mice received an intraperitoneal (i.p.) injection of 2.5 mg/kg deptropine three times a week for 3 weeks. ( a ) The tumor volume was measured every 2 or 3 days, and ( b ) the tumor weight was measured at the end of the experiment. Values were obtained from seven or eight samples, and results are presented as the mean ± standard error. * p < 0.05 vs. the control.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: Injection, Control

Journal: Cancers

Article Title: The Antihistamine Deptropine Induces Hepatoma Cell Death through Blocking Autophagosome-Lysosome Fusion

doi: 10.3390/cancers12061610

Figure Lengend Snippet: The possible mechanisms of deptropine-induced cell death in human hepatoma cells.

Article Snippet: Azatadine was obtained from The United States Pharmacopeial Convention (USP; Rockville, MD, USA) and deptropine citrate from the European Directorate for the Quality of Medicines (EDQM; Strasbourg, France).

Techniques: