Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 displays potent antiproliferative activity against pancreatic cancer cell lines. a The growth inhibition ratio in Aspc-1 cells after 6 days of treatment with four serial concentrations of various inhibitors as indicated. Red to blue, cell growth inhibition ratio from 100% to 0%. b A panel of human pancreatic adenocarcinoma cells were treated with CUDC-907 at gradient concentrations for 3 days, and the half-maximal inhibitory concentration (IC50) was measured using the SRB assay. Error bars represent the mean ± SD from three independent experiments. c Dose-dependent proliferation curves of Aspc-1, PANC-1, and Capan-1 cells over time. Cells were treated with CUDC-907 at gradient concentrations for 0, 1, 2, 3, or 4 days, and the relative cell growth ratio (%) was calculated using SoftMax Pro. Error bars represent the mean ± SD from three independent experiments. d Colony-formation assays. Aspc-1, PANC-1, and Capan-1 cells were measured with crystal violet staining after treatment with CUDC-907 at 5 to 20 nM for 14 days. e–g CUDC-907 arrests pancreatic cancer cells in the G2/M phase and induces apoptosis. Aspc-1 and Capan-1 cells were incubated with various concentrations (as indicated) of CUDC-907 for 24 or 48 h, and the percentage of each cell cycle phase (e) and apoptosis (f) was assessed by flow cytometry; (g) Aspc-1 and Capan-1 cells were collected for Western blotting with the indicated antibodies after treatment with CUDC-907 for 48 h

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: Activity Assay, Inhibition, Concentration Assay, Sulforhodamine B Assay, Staining, Incubation, Flow Cytometry, Western Blot

Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 downregulates c-Myc protein to inhibit cell proliferation. a c-Myc expression in pancreatic cancer cell lines compared with the normal cell line H6C7 evaluated by Western blotting. b c-Myc protein level alteration. Protein levels in Aspc-1, PANC-1, and Capan-1 cells treated with CUDC-907 (0, 25, 50, 100, and 500 nM) for 24 h were measured. The cell lysates were subjected to Western blotting analysis using the indicated antibodies. c c-Myc mRNA level alteration. Cells pretreated as described in b followed by qRT-PCR analysis of c-Myc level. Error bars represent the mean ± SD from three independent experiments. d Cell growth assay. Pancreatic cancer cells were transfected with c-Myc-specific siRNAs for 24 h and then transferred to a 96-well plate to be cultured for 3 or 6 days or to a 6-well plate for 14 days to form colonies. Relative cell proliferation rates were calculated by OD values, and colonies were scanned for crystal violet staining. Interference efficiency was verified by Western blotting. e Aspc-1-MYC and PANC-1-MYC cells, which were stably transfected with a mock or MYC-overexpression plasmid, were transfected with siRNAs against c-Myc or scramble for 24 h and then were transferred to 96-well plates to be cultured for 6 days, and the relative cell proliferation rate was calculated. Overexpression efficiency was verified by Western blotting. Error bars represent the mean ± SD from triplicates or three independent experiments. ***P < 0.001

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Growth Assay, Transfection, Cell Culture, Staining, Stable Transfection, Over Expression, Plasmid Preparation

Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 inhibits both the PI3K pathway and HDACs to suppress c-Myc function. a Individual PI3K or HDACs inhibitors suppress c-Myc. Aspc-1, PANC-1, and Capan-1 cells were treated with the indicated concentrations of the PI3K inhibitor GDC-0941 or the HDAC inhibitor vorinostat for 24 h, and the expression of c-Myc, phosphorylation of AKT and the acetylation of histone proteins were detected by Western blotting. b Synergistic effect of PI3K and HDAC inhibitors. IC50 values of vorinostat and GDC-0941 alone or in combination against Aspc-1, PANC-1, and Capan-1 cells were tested, and the combination index (CI) was calculated using the CompuSyn software. CI < 1, synergic effect; CI = 1, additive effect; CI > 1, antagonistic effect. Three independent experiments were exhibited. Error bars represent the mean ± SD from three independent experiments

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: Expressing, Phospho-proteomics, Western Blot, Software

Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 inhibits the PI3K pathway and leads to degradation of c-Myc. a PI3K pathway inhibition upregulates phosphor-c-Myc (Thr58) protein level. Aspc-1 and PANC-1 cells were treated with CUDC-907 or the PI3K inhibitor GDC-0941 for 2 h, and phosphor-c-Myc (Thr58) and total c-Myc protein levels were detected by Western blotting analysis using the indicated antibodies. Relative accumulation of phosphor-c-Myc (Thr58) to total c-Myc (normalized by the same Actin using the Image J software) was shown. b Aspc-1 and PANC-1 cells were treated with CUDC-907 (0, 25, 50, 100, and 500 nM) with (right) or without (left) proteasome inhibitor MG-132 for 2 h and then the cell lysates were subjected to Western blotting analysis to detect the c-Myc protein level

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: Inhibition, Western Blot, Software

Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 regulates c-Myc expression and cell proliferation through HDAC6 inhibition. a HDAC6 interference inhibits c-Myc protein levels. Aspc-1 and Capan-1 cells were transfected with individual siRNA against HDAC1, 2, 3, 4, and 6. HDAC6 interference inhibits c-Myc protein levels, as detected by Western blotting analysis using the indicated antibodies. b HDAC6 interference suppresses cell proliferation. Interference was performed as shown in Fig. 4a, and cells were transferred to a 96-well plate to be cultured for 3 days. The relative cell proliferation rate was calculated by OD values. Error bars represent the mean ± SD from three independent experiments. c HDAC6 reverses antiproliferation effect induced by CUDC-907 partially. Aspc-1-MOCK and Aspc-1-HDAC6 cells, which were stably transfected with a MOCK or HDAC6-overexpressing plasmid, were treated with CUDC-907 (0, 2, 5 nM) for 3 days, and the relative cell proliferation rate was calculated using SoftMax Pro. Error bars represent the mean ± SD from three independent experiments. d, e HDAC6 regulates c-Myc through the HDAC6–FOXO1–c-Myc axis. Aspc-1 cells were transfected with individual siRNA against HDAC6, FOXO1, or HDAC6-overexpressing plasmid, and the protein levels were detected by Western blotting analysis using the indicated antibodies (d). Also, a series of concentration of CUDC-907 were treated in Aspc-1, PANC-1, and Capan-1 cells for 24 h, and the FOXO1 and c-Myc protein levels were evaluated by Western blotting (e)

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: Expressing, Inhibition, Transfection, Western Blot, Cell Culture, Stable Transfection, Plasmid Preparation, Concentration Assay

Journal: Acta Pharmacologica Sinica

Article Title: CUDC-907 displays potent antitumor activity against human pancreatic adenocarcinoma in vitro and in vivo through inhibition of HDAC6 to downregulate c-Myc expression

doi: 10.1038/s41401-018-0108-5

Figure Lengend Snippet: CUDC-907 inhibits pancreatic cancer growth in vivo. a Tumor growth inhibition upon CUDC-907 treatment in Aspc-1 pancreatic xenografts. The curve of relative tumor volume after 20 days of treatment with CUDC-907 or vehicle and the corresponding curve of body weight are shown. Error bars represent the mean ± SD from seven individuals in each group. **P < 0.01. b Tumor morphology and weight. After 20 days, all nude mice were sacrificed, and the tumors were resected to be imaged and weighed. c Protein levels' alteration. Equal amounts of proteins from tumor tissues were evaluated for the phosphorylation of AKT and Erk and the acetylation of histone. d Immunohistochemistry staining. (i) Representative images of Ki67 and c-Myc staining of untreated xenografts and xenografts treated with CUDC-907. Scale bar, 100 μm. (ii) Quantification of immunohistochemistry staining was calculated by quick score (QS) method. Ten random areas from three individually stained tumors were captured. Error bars represent the mean ± SD from three independent experiments. **P < 0.01

Article Snippet: Compounds CUDC-907 was purchased from Med Chem Express (MCE) (NJ, USA), dissolved in dimethylsulfoxide at 10 mg/mL to form a stock solution and stored at −80 °C for in vitro studies.

Techniques: In Vivo, Inhibition, Phospho-proteomics, Immunohistochemistry, Staining

Journal: International Journal of Oncology

Article Title: The dual HDAC and PI3K inhibitor, CUDC-907, inhibits tumor growth and stem-like properties by suppressing PTX3 in neuroblastoma

doi: 10.3892/ijo.2023.5602

Figure Lengend Snippet: CUDC-907 inhibits the proliferation, induces the apoptosis, and reduces the migratory ability of NB cells. (A and B) The indicated NB cell lines were treated with various concentrations of CUDC-907, and the IC50 value was calculated at 72 h. Data are presented as the mean ± SD. (C) Effect of various concentrations of CUDC-907 on the colony formation of SK-N-SH, SK-N-BE and IMR32 cells. The histogram on the right indicates the number of clones. **P<0.01, *** P<0.001 and **** P<0.0001, vs. NC; ns, not significant. (D) SK-N-SH, SK-N-BE and IMR32 cells were collected for western blot analysis with the indicated antibodies against apoptosis-related proteins following treatment with various concentrations of CUDC-907 for 24 h. NB, neuroblastoma; NC, negative control.

Article Snippet: CUDC-907 was purchased from TargetMol.

Techniques: Colony Assay, Clone Assay, Western Blot, Analysis, Negative Control, Cell Viability Assay

Journal: International Journal of Oncology

Article Title: The dual HDAC and PI3K inhibitor, CUDC-907, inhibits tumor growth and stem-like properties by suppressing PTX3 in neuroblastoma

doi: 10.3892/ijo.2023.5602

Figure Lengend Snippet: CUDC-907 inhibits the growth of NB xenografts in vivo . (A) Schematic diagram illustrating the administration mode and time in mice. The curves of (B) relative tumor volume and (C) body weight of the mice treated with CUDC-907 or the vehicle. Error bars represent the mean ± SD in each group. *** P<0.001. Comparison of (D) tumor size and (E) tumor weight between the vehicle and CUDC-907-treated group. ** P<0.01. (F) Changes in the protein levels of H3, H3K27ac, AKT and p-AKT in the animal tissues between the vehicle and CUDC-907-treated group. (G and H) Changes in Ki67 expression in NB tissues between the vehicle and CUDC-907-treated group. ** P<0.01. NB, neuroblastoma; H3, histone H3.

Article Snippet: CUDC-907 was purchased from TargetMol.

Techniques: In Vivo, Comparison, Expressing, Xenograft Assay, Western Blot, Immunohistochemistry

Journal: International Journal of Oncology

Article Title: The dual HDAC and PI3K inhibitor, CUDC-907, inhibits tumor growth and stem-like properties by suppressing PTX3 in neuroblastoma

doi: 10.3892/ijo.2023.5602

Figure Lengend Snippet: CUDC-907 downregulates MYCN expression, and suppresses the activation of the PI3K/AKT and MAPK/ERK pathways in NB cells. (A) Western blot analysis and (B) reverse transcription-quantitative PCR of MYCN expression in the indicated NB cells. (C and D) Western blot analysis of MYCN expression in SK-N-BE cells treated (C) with various concentrations of CUDC-907 for 24 h or (D) with a fixed concentration of CUDC-907 (50 nM) for the indicated periods of time. (E and F) Western blot analysis of MYCN expression in IMR32 cells treated (E) with various concentrations of CUDC-907 for 24 h or (F) with a fixed concentration of CUDC-907 (50 nM) for the indicated periods of time. (G) Western blot analysis of H3K27ac, AKT, p-AKT, MEK, p-MEK, ERK and p-ERK expression in the SK-N-SH, SK-N-BE and IMR32 cells treated with various concentrations of CUDC-907 for 24 h. NB, neuroblastoma; H3, histone H3.

Article Snippet: CUDC-907 was purchased from TargetMol.

Techniques: Expressing, Activation Assay, Western Blot, Analysis, Reverse Transcription, Real-time Polymerase Chain Reaction, Concentration Assay

Journal: International Journal of Oncology

Article Title: The dual HDAC and PI3K inhibitor, CUDC-907, inhibits tumor growth and stem-like properties by suppressing PTX3 in neuroblastoma

doi: 10.3892/ijo.2023.5602

Figure Lengend Snippet: NB cells treated with CUDC-907 exhibit weaker CSC-like properties. (A) Sphere images of the first, second and third generation of SK-N-BE cells treated with various concentrations of CUDC-907. (B) The sphere numbers of the first, second and third generation of SK-N-BE cells treated with various concentrations of CUDC-907. * P<0.05, ** P<0.01 and *** P<0.001; ns, not significant. (C) Reverse transcription-quantitative PCR of the expression of the stem cell markers, CD44, SOX2, OCT4 and BMI-1, in the first, second and third generation of SK-N-BE cells treated with the indicated concentrations of CUDC-907. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001, vs. NC group; ns, not significant. (D) Following CUDC-907 treatment, western blot analyses were performed on the indicated stem cell markers in SK-N-BE cells at the protein level. NB, neuroblastoma; SOX2, sex determining region Y-box 2; OCT4, octamer-binding transcription factor 4; BMI-1, B-cell-specific Moloney murine leukemia virus integration site 1.

Article Snippet: CUDC-907 was purchased from TargetMol.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Stem Cell, Western Blot, Binding Assay, Virus

Journal: International Journal of Oncology

Article Title: The dual HDAC and PI3K inhibitor, CUDC-907, inhibits tumor growth and stem-like properties by suppressing PTX3 in neuroblastoma

doi: 10.3892/ijo.2023.5602

Figure Lengend Snippet: CUDC-907 inhibits the stem-like properties of NB by suppressing PTX3. (A) mRNA levels of PTX3 in cells treated with 50 nM CUDC-907 for 24 h. *** P<0.001, vs. DMSO control. (B) mRNA levels of PTX3 in cells transfected with PTX3 siRNA. *** P<0.001 and **** P<0.0001, vs. negative control. (C) mRNA levels of PTX3, CD44, MYCN, BMI-1, SOX2 and OCT4 expression in cells transfected with negative control or PTX3 siRNA. * P<0.05 and ** P<0.01, vs. negative control. (D) PTX3, CD44 and p-AKT protein expression levels in cells transfected with PTX3 siRNA. (E) Sphere formation of NB cells transfected with PTX3 siRNA. * P<0.05, vs. negative control. (F) Sphere formation of empty vector-transfected cells exposed to DMSO, empty vector-transfected cells exposed to 50 nM CUDC-907 and PTX3-overexpressing cells exposed to 50 nM CUDC-907. * P<0.05 and ** P<0.01; ns, not significant. (G-I) The mRNA/protein expression levels of PTX3 and CD44 in empty vector-transfected cells exposed to DMSO, empty vector-transfected cells exposed to 50 nM CUDC-907 and PTX3-overexpressing cells exposed to 50 nM CUDC-907. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001. NB, neuroblastoma; PTX3, pentraxin 3; SOX2, sex determining region Y-box 2; OCT4, octamer-binding transcription factor 4; BMI-1, B-cell-specific Moloney murine leukemia virus integration site 1.

Article Snippet: CUDC-907 was purchased from TargetMol.

Techniques: Control, Transfection, Negative Control, Expressing, Plasmid Preparation, Binding Assay, Virus, Real-time Polymerase Chain Reaction, Western Blot