Journal: World Journal of Gastroenterology

Article Title: Wogonin derivative V8 enhances bortezomib efficacy in gastric carcinoma by disrupting lysosome-mediated drug resistance

doi: 10.3748/wjg.v32.i8.113299

Figure Lengend Snippet: Lysosome affects the anticancer effect of bortezomib. A: Intracellular drug concentration was examined using liquid chromatography/mass spectrometry after cells were treated with bortezomib (BTZ) (100 nM) and bafilomycin A1 (100 nM) alone or in combination for 6 hours; B and C: MGC-803 cells were treated with indicated concentration of BTZ and L-leucyl-L-leucine methyl ester hydrochloride (1 mmol/L) for 6 hours, followed by staining of LysoSensor Green DND-189. Representative images (B) and quantification data (C) of flow cytometry were shown; D and E: Representative images (D) and quantified results (E) of immunofluorescent analysis for LAMP1 expression in different gastric cancer cell lines (scale bar: 10 μm); F: Cell viability assay was performed in different gastric cancer cells treated with BTZ; G: Spearman correlation was determined between half-maximal inhibitory concentration value for BTZ and cellular LAMP1 expression. b P < 0.01. c P < 0.001. d P < 0.0001. BTZ: Bortezomib; Baf-A1: Bafilomycin A1; LLOMe: L-leucyl-L-leucine methyl ester hydrochloride; DMSO: Dimethyl sulfoxide; MFI: Mean fluorescence intensity; DAPI: 4’,6-Diamidino-2-phenylindole; NS: No significance; IC 50 : Half-maximal inhibitory concentration.

Article Snippet: Cells treated with V8 and BTZ alone or combination were harvested and stained with the Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (A211-01/02; Vazyme, Nanjing, Jiangsu Province, China), LysoSensor TM Green DND-189 (40767ES50; Yeasen Biotechnology).

Techniques: Concentration Assay, Liquid Chromatography, Mass Spectrometry, Staining, Flow Cytometry, Expressing, Viability Assay, Fluorescence

Journal: World Journal of Gastroenterology

Article Title: Wogonin derivative V8 enhances bortezomib efficacy in gastric carcinoma by disrupting lysosome-mediated drug resistance

doi: 10.3748/wjg.v32.i8.113299

Figure Lengend Snippet: V8 induces lysosomal cell death in gastric cancer cells. A: Structure of synthetic flavonoid derivate V8; B and C: Gastric cancer cells were treated with V8 at indicated concentration for 24 hours. Representative images (B) and quantified data (C) for cell death are shown; D and E: MGC-803 cells were treated with V8 and L-leucyl-L-leucine methyl ester hydrochloride (1 mmol/L) for 6 hours, followed by staining with LysoSensor Green DND-189 dye. Representative images (D) and quantification data (E) are shown; F and G: MGC-803 cells were treated with V8 (9 μM), followed by staining with LysoTracker red dye. Representative images and quantified intensity of immunofluorescence analysis are shown (scale bar: 25 μm); H-K: MGC-803 cells were transfected with small interfering RNA against CSTB gene or treated with bafilomycin A1 (100 nM), followed by V8 (12 μM) for 24 hours. Representative images (H and J) and quantification data (I and K) of flow cytometry analysis for cell death are shown. a P < 0.05. b P < 0.01. c P < 0.001. d P < 0.0001. V-FITC: Annexin V-fluorescein isothiocyanate; PI: Propidium iodide; LLOMe: L-leucyl-L-leucine methyl ester hydrochloride; DMSO: Dimethyl sulfoxide; MFI: Mean fluorescence intensity; DAPI: 4’,6-Diamidino-2-phenylindole; NS: No significance; Baf-A1: Bafilomycin A1; siCtrl: Negative control small interfering RNA; si CTSB : Small interfering RNA targeting CTSB .

Article Snippet: Cells treated with V8 and BTZ alone or combination were harvested and stained with the Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (A211-01/02; Vazyme, Nanjing, Jiangsu Province, China), LysoSensor TM Green DND-189 (40767ES50; Yeasen Biotechnology).

Techniques: Concentration Assay, Staining, Immunofluorescence, Transfection, Small Interfering RNA, Flow Cytometry, Fluorescence, Negative Control