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cct196969  (MedChemExpress)
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MedChemExpress cct196969
<t>CCT196969</t> significantly inhibits TNBC in vitro and in vivo. A - G . 4T1 cells and MDA-MB-231 cells were treated with different concentrations of CCT196969 for 48 h, and the following experiments were performed: A . CCK-8 experiment, with data from three repeated experiments. B - C . Clone formation experiment was repeated three times, the number of cell colonies was counted, and the effect of CCT196969 on cell growth was observed. Figure B is the representative result, and Figure C is the statistical figure. D - E . Cell invasion and migration experiment, three independent data were collected for statistical analysis. Figure D is the typical result, and Figure E is the statistical figure. Scale bars, 20 μm. F - G . Apoptosis experiment, Annexin V-FITC/PI double staining flow assay was used to collect three independent data for statistical analysis. Figure F represents the typical result and Figure G is the statistical graph. H . Comparison of tumor tissue size between Control group and CCT196969 group (10 mg/kg, gavage once a day), where n = 6 in Control group and n = 6 in CCT196969 group. I . Tumor weight statistics of mice in Control group and CCT196969 treatment group. J . Tumor growth curves of mice in Control group and CCT196969 treatment group. K . Weight changes of mice in Control group and CCT196969 treatment group. All values are mean ± SD. The statistical significance was determined by t test, *, P < 0.05; **, P < 0.01; ***, P < 0.001
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CCT196969 significantly inhibits TNBC in vitro and in vivo. A - G . 4T1 cells and MDA-MB-231 cells were treated with different concentrations of CCT196969 for 48 h, and the following experiments were performed: A . CCK-8 experiment, with data from three repeated experiments. B - C . Clone formation experiment was repeated three times, the number of cell colonies was counted, and the effect of CCT196969 on cell growth was observed. Figure B is the representative result, and Figure C is the statistical figure. D - E . Cell invasion and migration experiment, three independent data were collected for statistical analysis. Figure D is the typical result, and Figure E is the statistical figure. Scale bars, 20 μm. F - G . Apoptosis experiment, Annexin V-FITC/PI double staining flow assay was used to collect three independent data for statistical analysis. Figure F represents the typical result and Figure G is the statistical graph. H . Comparison of tumor tissue size between Control group and CCT196969 group (10 mg/kg, gavage once a day), where n = 6 in Control group and n = 6 in CCT196969 group. I . Tumor weight statistics of mice in Control group and CCT196969 treatment group. J . Tumor growth curves of mice in Control group and CCT196969 treatment group. K . Weight changes of mice in Control group and CCT196969 treatment group. All values are mean ± SD. The statistical significance was determined by t test, *, P < 0.05; **, P < 0.01; ***, P < 0.001

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCT196969 inhibits TNBC by targeting the HDAC5/RXRA/ASNS axis to down-regulate asparagine synthesis

doi: 10.1186/s13046-025-03494-5

Figure Lengend Snippet: CCT196969 significantly inhibits TNBC in vitro and in vivo. A - G . 4T1 cells and MDA-MB-231 cells were treated with different concentrations of CCT196969 for 48 h, and the following experiments were performed: A . CCK-8 experiment, with data from three repeated experiments. B - C . Clone formation experiment was repeated three times, the number of cell colonies was counted, and the effect of CCT196969 on cell growth was observed. Figure B is the representative result, and Figure C is the statistical figure. D - E . Cell invasion and migration experiment, three independent data were collected for statistical analysis. Figure D is the typical result, and Figure E is the statistical figure. Scale bars, 20 μm. F - G . Apoptosis experiment, Annexin V-FITC/PI double staining flow assay was used to collect three independent data for statistical analysis. Figure F represents the typical result and Figure G is the statistical graph. H . Comparison of tumor tissue size between Control group and CCT196969 group (10 mg/kg, gavage once a day), where n = 6 in Control group and n = 6 in CCT196969 group. I . Tumor weight statistics of mice in Control group and CCT196969 treatment group. J . Tumor growth curves of mice in Control group and CCT196969 treatment group. K . Weight changes of mice in Control group and CCT196969 treatment group. All values are mean ± SD. The statistical significance was determined by t test, *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: For combined treatment experiment, the experimental group was given 5 mg/kg CCT196969 or 40UI ASNase (MCE, HY-P1923) every day, the combined group was given 5 mg/kg CCT196969 and 40UI ASNase every day, and the control group was given the same amount of sterile water.

Techniques: In Vitro, In Vivo, CCK-8 Assay, Migration, Double Staining, Comparison, Control

CCT196969 inhibits TNBC by down-regulating ASNS to inhibit asparagine synthesis and its downstream mTORC pathway. A . The volcano map showed 2614 differential genes (|log2Fold Change|≥1, FDR < 0.05), with up-regulated genes in red markers, down-regulated genes in green markers, and no significant differential genes in blue markers. B . KEGG enrichment analysis showed the 20 pathways with the most significant differences. C . GSEA enrichment map of amino acid starvation. D . Differential metabolite clustering heat map (Fold_Change ≥ 2, Fold_Change ≤ 0.5 or VIP ≥ 1). E . Radar map of the top ten metabolites with the largest FC values. F . Bubble map of combined transcriptome and metabolome analysis. G . The top three of the co-enrichment pathway sorted by P-value. H . qPCR was used to detect the mRNA expression of ASNS in 4T1 cells treated with different concentrations of CCT196969 for 48 h. I . Western blot analysis of ASNS protein expression in 4T1 cells treated with different concentrations of CCT196969 for 48 h. J . Western blot detection of ASNS protein expression in tumor tissues of mice in Control group (N) and CCT196969 treatment group (T). K . The content of asparagine in blood of mice was analyzed by mass spectrometry. L . Western blot analysis of mTORC1 pathway protein expression in 4T1 cells treated with different concentrations of CCT196969 for 48 h. M . Western blot analysis of mTORC1 pathway protein expression in mouse tumor tissues. The samples used for J , K and M are from Fig. . All membranes were cut before antibody incubation, and all the following Western blotting figure legends are the same as this one. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCT196969 inhibits TNBC by targeting the HDAC5/RXRA/ASNS axis to down-regulate asparagine synthesis

doi: 10.1186/s13046-025-03494-5

Figure Lengend Snippet: CCT196969 inhibits TNBC by down-regulating ASNS to inhibit asparagine synthesis and its downstream mTORC pathway. A . The volcano map showed 2614 differential genes (|log2Fold Change|≥1, FDR < 0.05), with up-regulated genes in red markers, down-regulated genes in green markers, and no significant differential genes in blue markers. B . KEGG enrichment analysis showed the 20 pathways with the most significant differences. C . GSEA enrichment map of amino acid starvation. D . Differential metabolite clustering heat map (Fold_Change ≥ 2, Fold_Change ≤ 0.5 or VIP ≥ 1). E . Radar map of the top ten metabolites with the largest FC values. F . Bubble map of combined transcriptome and metabolome analysis. G . The top three of the co-enrichment pathway sorted by P-value. H . qPCR was used to detect the mRNA expression of ASNS in 4T1 cells treated with different concentrations of CCT196969 for 48 h. I . Western blot analysis of ASNS protein expression in 4T1 cells treated with different concentrations of CCT196969 for 48 h. J . Western blot detection of ASNS protein expression in tumor tissues of mice in Control group (N) and CCT196969 treatment group (T). K . The content of asparagine in blood of mice was analyzed by mass spectrometry. L . Western blot analysis of mTORC1 pathway protein expression in 4T1 cells treated with different concentrations of CCT196969 for 48 h. M . Western blot analysis of mTORC1 pathway protein expression in mouse tumor tissues. The samples used for J , K and M are from Fig. . All membranes were cut before antibody incubation, and all the following Western blotting figure legends are the same as this one. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: For combined treatment experiment, the experimental group was given 5 mg/kg CCT196969 or 40UI ASNase (MCE, HY-P1923) every day, the combined group was given 5 mg/kg CCT196969 and 40UI ASNase every day, and the control group was given the same amount of sterile water.

Techniques: Expressing, Western Blot, Control, Mass Spectrometry, Incubation

CCT196969 inhibits ASNS expression by up-regulating RXRA. A . 4T1 cells treated with CCT196969 for 48 h were then treated with CHX, and the protein was collected after 0, 12, 24, 36 h, respectively. Western blot analysis was performed to detect the degradation of ASNS protein (3 repetitions, the figure above is representative and the figure below is statistical). B . Statistical maps of proteomic analysis between control group and CCT196969 treatment group. C . Differential protein volcano map (red: significantly up-regulated, blue: significantly down-regulated, gray: no significant difference). D . Venn diagram of transcription factors for combined analysis of proteome and transcriptome. E . Western blot analysis of ASNS protein expression after POU2F1 knockdown. F . Western blot analysis of ASNS protein expression after KLF5 knockdown. G . Western blot analysis of ASNS protein expression after RXRA knockdown. H . qPCR was used to detect ASNS mRNA expression after RXRA knockdown. I . Western blot analysis of RXRA and ASNS protein expression after CCT196969 treatment. J . mRNA expression of RXRA and ASNS after CCT196969 treatment was detected by qPCR. K - L . Western blot analysis of ASNS protein expression after CCT196969 treatment in sgNC-4T1 and sgRXRA-4T1 cells ( K ), shNC-4T1 and shRXRA-4T1 cells ( L ). M - O . shNC-4T1 and shRXRA-4T1 cells were treated or not treated with CCT196969, and the following experiments were performed: M . CCK-8 experiment (3 independent experiments). N . clone formation experiment (3 repetitions, representative graph on the left, statistical graph on the right). O . Transwell experiment (3 independent experiments, representative graph on the left and statistical graph on the right). Scale bars, 20 μm. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCT196969 inhibits TNBC by targeting the HDAC5/RXRA/ASNS axis to down-regulate asparagine synthesis

doi: 10.1186/s13046-025-03494-5

Figure Lengend Snippet: CCT196969 inhibits ASNS expression by up-regulating RXRA. A . 4T1 cells treated with CCT196969 for 48 h were then treated with CHX, and the protein was collected after 0, 12, 24, 36 h, respectively. Western blot analysis was performed to detect the degradation of ASNS protein (3 repetitions, the figure above is representative and the figure below is statistical). B . Statistical maps of proteomic analysis between control group and CCT196969 treatment group. C . Differential protein volcano map (red: significantly up-regulated, blue: significantly down-regulated, gray: no significant difference). D . Venn diagram of transcription factors for combined analysis of proteome and transcriptome. E . Western blot analysis of ASNS protein expression after POU2F1 knockdown. F . Western blot analysis of ASNS protein expression after KLF5 knockdown. G . Western blot analysis of ASNS protein expression after RXRA knockdown. H . qPCR was used to detect ASNS mRNA expression after RXRA knockdown. I . Western blot analysis of RXRA and ASNS protein expression after CCT196969 treatment. J . mRNA expression of RXRA and ASNS after CCT196969 treatment was detected by qPCR. K - L . Western blot analysis of ASNS protein expression after CCT196969 treatment in sgNC-4T1 and sgRXRA-4T1 cells ( K ), shNC-4T1 and shRXRA-4T1 cells ( L ). M - O . shNC-4T1 and shRXRA-4T1 cells were treated or not treated with CCT196969, and the following experiments were performed: M . CCK-8 experiment (3 independent experiments). N . clone formation experiment (3 repetitions, representative graph on the left, statistical graph on the right). O . Transwell experiment (3 independent experiments, representative graph on the left and statistical graph on the right). Scale bars, 20 μm. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: For combined treatment experiment, the experimental group was given 5 mg/kg CCT196969 or 40UI ASNase (MCE, HY-P1923) every day, the combined group was given 5 mg/kg CCT196969 and 40UI ASNase every day, and the control group was given the same amount of sterile water.

Techniques: Expressing, Western Blot, Control, Knockdown, CCK-8 Assay

CCT196969 up-regulates RXRA by targetly inhibiting HDAC5, and then down-regulates ASNS expression, inhibiting TNBC. A . CCT196969 was fixed with 3D optical crosslinking chip, 4T1 cell lysate was circulated, targeted proteins were identified by LC-MS, and heat maps were generated by clustering according to Score and Relative Quantity. Western blot was used to detect the ASNS protein expression after BRAF knockdown ( B )/RAF1 knockdown ( C )/SRC knockdown ( D ), the thermostability of HDAC5 ( E )/HDAC5 protein expression ( F ) before and after CCT196969 treatment and the protein expression of RXRA and ASNS after HDAC5 knockdown ( G ). H - J . shNC-4T1 and shHDAC5-4T1 cells were treated or not treated with CCT196969, and the following experiments were performed: H . CCK-8 experiment (3 independent experiments). I . Clone formation experiment (3 repetitions, representative graph on the left, statistical graph on the right). J . Transwell experiment (3 independent experiments, representative graph on the left, statistical graph on the right). Scale bars, 20 μm. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCT196969 inhibits TNBC by targeting the HDAC5/RXRA/ASNS axis to down-regulate asparagine synthesis

doi: 10.1186/s13046-025-03494-5

Figure Lengend Snippet: CCT196969 up-regulates RXRA by targetly inhibiting HDAC5, and then down-regulates ASNS expression, inhibiting TNBC. A . CCT196969 was fixed with 3D optical crosslinking chip, 4T1 cell lysate was circulated, targeted proteins were identified by LC-MS, and heat maps were generated by clustering according to Score and Relative Quantity. Western blot was used to detect the ASNS protein expression after BRAF knockdown ( B )/RAF1 knockdown ( C )/SRC knockdown ( D ), the thermostability of HDAC5 ( E )/HDAC5 protein expression ( F ) before and after CCT196969 treatment and the protein expression of RXRA and ASNS after HDAC5 knockdown ( G ). H - J . shNC-4T1 and shHDAC5-4T1 cells were treated or not treated with CCT196969, and the following experiments were performed: H . CCK-8 experiment (3 independent experiments). I . Clone formation experiment (3 repetitions, representative graph on the left, statistical graph on the right). J . Transwell experiment (3 independent experiments, representative graph on the left, statistical graph on the right). Scale bars, 20 μm. Data were mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: For combined treatment experiment, the experimental group was given 5 mg/kg CCT196969 or 40UI ASNase (MCE, HY-P1923) every day, the combined group was given 5 mg/kg CCT196969 and 40UI ASNase every day, and the control group was given the same amount of sterile water.

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Generated, Western Blot, Knockdown, CCK-8 Assay

There is a positive correlation between the expression of HDAC5 and ASNS. A . The correlation between HDAC5 and ASNS mRNA levels was analyzed by TCGA database. B . Immunohistochemical representation of HDAC5 and ASNS in cancer tissue chips of 80 TNBC patients. C . Correlation between HDAC5 expression and ASNS expression in 80 TNBC patients. D - G . TCGA database was used to analyze the relationship between HDAC5 expression level and clinical prognosis (OS) in BC patients ( D ), ER-positive BC patients ( E ), HER2-positive BC patients ( F ) and TNBC patients ( G ). H. Molecular mechanism diagram, CCT196969 down-regulates asparagine synthesis by targeting the HDAC5/RXRA/ASNS axis and further inhibits the mTORC pathway. Ultimately, CCT196969 inhibited the proliferation, invasion and migration of triple-negative breast cancer

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCT196969 inhibits TNBC by targeting the HDAC5/RXRA/ASNS axis to down-regulate asparagine synthesis

doi: 10.1186/s13046-025-03494-5

Figure Lengend Snippet: There is a positive correlation between the expression of HDAC5 and ASNS. A . The correlation between HDAC5 and ASNS mRNA levels was analyzed by TCGA database. B . Immunohistochemical representation of HDAC5 and ASNS in cancer tissue chips of 80 TNBC patients. C . Correlation between HDAC5 expression and ASNS expression in 80 TNBC patients. D - G . TCGA database was used to analyze the relationship between HDAC5 expression level and clinical prognosis (OS) in BC patients ( D ), ER-positive BC patients ( E ), HER2-positive BC patients ( F ) and TNBC patients ( G ). H. Molecular mechanism diagram, CCT196969 down-regulates asparagine synthesis by targeting the HDAC5/RXRA/ASNS axis and further inhibits the mTORC pathway. Ultimately, CCT196969 inhibited the proliferation, invasion and migration of triple-negative breast cancer

Article Snippet: For combined treatment experiment, the experimental group was given 5 mg/kg CCT196969 or 40UI ASNase (MCE, HY-P1923) every day, the combined group was given 5 mg/kg CCT196969 and 40UI ASNase every day, and the control group was given the same amount of sterile water.

Techniques: Expressing, Immunohistochemical staining, Migration