Journal: Oncology Letters

Article Title: Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells

doi: 10.3892/ol.2019.10664

Figure Lengend Snippet: Viability of HCT116 cells investigated by MTT assay. (A) Structure of MHY695. (B) HCT116 cells were treated with vehicle or MHY695 for 24 h at the indicated concentrations. *P<0.05 and ***P<0.001 vs. the vehicle-treated group. MHY695, (Z)-5-(3-ethoxy-4-hydroxybenzylidene)-2- thioxothiazolidin-4-one.

Article Snippet: A suspension of 3-ethoxy-4-hydroxybenzaldehyde (purity, 99%; 300 mg; 1.81 mmol; Sigma-Aldrich; Merck KGaA) and rhodanine (purity, 97%; 2-thioxothiazolidin-4-one; 264.5 mg; 1.99 mmol; Sigma-Aldrich; Merck KGaA) in 4 ml ethanol was refluxed for 5 h in the presence of piperidine (purity, 99%; 0.05 ml; 0.54 mmol; Sigma-Aldrich; Merck KGaA) ( ).

Techniques: MTT Assay

Journal: Oncology Letters

Article Title: Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells

doi: 10.3892/ol.2019.10664

Figure Lengend Snippet: MHY695 induces cytotoxicity by promoting apoptosis. (A) Morphological changes in HCT116 colon cancer cells were observed by confocal microscopy. Cells were treated with vehicle or MHY695 (25 or 50 µM) for 24 h. Apoptotic body formation (red arrows) was observed using DAPI staining (blue) in the MHY695-treated groups. The release of cytochrome C (green) was also observed in the MHY695-treated groups. (B) Apoptotic or necrotic cells were identified using flow cytometry. HCT116 cells were treated with MHY695 or vehicle for 24 h, prior to staining with Annexin V and PI. Annexin V-bound or PI-positive cells were subsequently quantified. MHY695, (Z)-5-(3-ethoxy-4-hydroxybenzylidene)-2-thioxothiazolidin-4-one; PI, propidium iodide.

Article Snippet: A suspension of 3-ethoxy-4-hydroxybenzaldehyde (purity, 99%; 300 mg; 1.81 mmol; Sigma-Aldrich; Merck KGaA) and rhodanine (purity, 97%; 2-thioxothiazolidin-4-one; 264.5 mg; 1.99 mmol; Sigma-Aldrich; Merck KGaA) in 4 ml ethanol was refluxed for 5 h in the presence of piperidine (purity, 99%; 0.05 ml; 0.54 mmol; Sigma-Aldrich; Merck KGaA) ( ).

Techniques: Confocal Microscopy, Staining, Flow Cytometry

Journal: Oncology Letters

Article Title: Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells

doi: 10.3892/ol.2019.10664

Figure Lengend Snippet: MHY695 modulates the expression of apoptosis-associated proteins. HCT116 cells were treated with vehicle or MHY695 (25 or 50 µM) for 24 h. Cellular proteins were separated using SDS-PAGE and transferred to PVDF membranes. The bands were detected using ECL solution and a chemiluminescence system. The bands are representative of three independent experiments. MHY695, (Z)-5-(3-ethoxy- 4-hydroxybenzylidene)-2-thioxothiazolidin-4-one; p-, phospho-; Akt, protein kinase B; Bad, Bcl-2-associated agonist of cell death; Mcl-1, induced myeloid leukaemia cell differentiation protein; PARP, poly(ADP-ribose) polymerase.

Article Snippet: A suspension of 3-ethoxy-4-hydroxybenzaldehyde (purity, 99%; 300 mg; 1.81 mmol; Sigma-Aldrich; Merck KGaA) and rhodanine (purity, 97%; 2-thioxothiazolidin-4-one; 264.5 mg; 1.99 mmol; Sigma-Aldrich; Merck KGaA) in 4 ml ethanol was refluxed for 5 h in the presence of piperidine (purity, 99%; 0.05 ml; 0.54 mmol; Sigma-Aldrich; Merck KGaA) ( ).

Techniques: Expressing, SDS Page, Cell Differentiation

Journal: Oncology Letters

Article Title: Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells

doi: 10.3892/ol.2019.10664

Figure Lengend Snippet: In vivo anti-cancer effects of MHY695 in a mouse xenograft model. (A) Tumors were resected from the mice following euthanasia. (B) Tumor size was measured daily with a caliper. Tumor volume (mm 3 ) was calculated as L 2 × S × 0.5. L, long diameter; S, short diameter. (C) Tumor weight was measured following resection. MHY695 (2 mg/kg) was subcutaneously injected into each tumor daily. *P<0.05, **P<0.01 and ***P<0.001 vs. the vehicle-treated group. (D) In vivo anti-cancer effects of MHY695 (5 mg/kg) were visualized using GFP-tagged HCT116 stem cells. MHY695, (Z)-5-(3-ethoxy-4-hydroxybenzylidene)-2-thioxothiazolidin-4-one.

Article Snippet: A suspension of 3-ethoxy-4-hydroxybenzaldehyde (purity, 99%; 300 mg; 1.81 mmol; Sigma-Aldrich; Merck KGaA) and rhodanine (purity, 97%; 2-thioxothiazolidin-4-one; 264.5 mg; 1.99 mmol; Sigma-Aldrich; Merck KGaA) in 4 ml ethanol was refluxed for 5 h in the presence of piperidine (purity, 99%; 0.05 ml; 0.54 mmol; Sigma-Aldrich; Merck KGaA) ( ).

Techniques: In Vivo, Injection

Journal: Oncology Letters

Article Title: Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells

doi: 10.3892/ol.2019.10664

Figure Lengend Snippet: Cytotoxic effects of MHY695 are independent of p53. (A) NCM460, HT-29, Caco-2, DLD-1 and HCT116 cells were treated with various concentrations of MHY695. *P<0.05, **P<0.01 and ***P<0.001 vs. the vehicle-treated group. (B) Expression levels of phospho-Akt and cleaved PARP were investigated in NCM460, Caco-2, DLD-1 and HT-29 cells by western blot analysis. Cells were treated with vehicle or MHY695 (25 or 50 µM) for 24 h. (C) Cell viability of p53 wt and p53 null HCT116 cells were investigated by MTT assay. p53 wt or p53 null HCT116 cells were treated with vehicle or MHY695 at the specified concentrations for 24 h. (D) Downstream proteins of the p53-associated apoptosis pathway were investigated by western blot analysis. p53 wt HCT116 cells were treated with vehicle or MHY695 at the specified concentrations for 24 h. MHY695, (Z)-5-(3-ethoxy-4-hydroxybenzylidene)-2-thioxothiazolidin-4-one; wt, wild-type; p-Akt, phospho-protein kinase B; PARP, poly(ADP-ribose) polymerase; PUMA, Bcl-2-binding component 3; Bax, Apoptosis regulator BAX; Bak, Bcl-2 homologous antagonist/killer.

Article Snippet: A suspension of 3-ethoxy-4-hydroxybenzaldehyde (purity, 99%; 300 mg; 1.81 mmol; Sigma-Aldrich; Merck KGaA) and rhodanine (purity, 97%; 2-thioxothiazolidin-4-one; 264.5 mg; 1.99 mmol; Sigma-Aldrich; Merck KGaA) in 4 ml ethanol was refluxed for 5 h in the presence of piperidine (purity, 99%; 0.05 ml; 0.54 mmol; Sigma-Aldrich; Merck KGaA) ( ).

Techniques: Expressing, Western Blot, MTT Assay, Binding Assay