Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Dose-response curves and IC₅₀ determination of CB2R compounds across gastric cancer cell lines. MTT assays were performed at 48 and 72 h to calculate IC₅₀ values. The graphs represented the results expressed as % of vitality at 48 h and displayed five coloured sigmoid curves, each corresponding to a different CB2R compound: blue for the antagonist AM630 , red for CC48 , green for Fi9 , purple for ASF151 and yellow for the reference compound 1 . The corresponding IC₅₀ values with their standard error of the mean (SEM), are shown alongside. Data are from one of three independent experiments; each performed in triplicate

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: CB2R-binding compounds counter PTX resistance and promote PTX-mediated inhibition of cell proliferation.The Ki67 cytofluorimetric assay was performed after treatment of PTX-sensitive HGC27 cells and their resistant counterparts and NCI-N87 cells with a combination of 10µM CC48 or Fi9 or ASF151 or 1 or 6 µM AM630 with 4 µM PTX. A ) Representative flow cytometry charts after exposure to CC48 and reporting the percentage of Ki67 negative (blue) and positive (red) cells; the Ki67 + population of the control is shown in grey. B ) Statistical charts reporting the results obtained in HGC27-S, HGC27-R and NCI-N87, from three independent experiments and expressed as means ± SD. Statistical analysis was assessed by comparing the values obtained using single drug treatment to those of corresponding untreated cells and the combined treatments to those with PTX alone, * p < 0.05; ** p < 0.01

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Binding Assay, Inhibition, Cytofluorimetric Assay, Flow Cytometry, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Western blot analysis of the expression and activation levels of key proteins involved in cell proliferation. Representative western blotting analyses performed in HGC27-S/R and statistical charts reporting the results from three independent experiments expressed as means ± SD. The expression of the phosphorylated and/or total forms of TSC2, AKT, p70, S6, 4EBP1, PI3K, and ERK1/2 after 48 h of treatment with the compounds AM630 , CC48 , Fi9 , ASF151 and 1 . The expression levels of each of the investigated proteins were normalized to the actin level, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Western Blot, Expressing, Activation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Western blot analysis of expression levels of key proteins involved in autophagy. A) Representative WB experiments showing the expression levels of autophagy-involved proteins ATG5, ATG7, ATG12, Beclin-1, LC3-II in PTX-sensitive and resistant HGC27 cell lines and AGS after 48 h of treatment with the compounds AM630 , CC48 , Fi9 , ASF151 and 1 . B ) Statistical charts reporting the results from three independent experiments and expressed as means ± SD. Expression levels of each of the investigated proteins were normalized to the Tubulin level, * p < 0.05; ** p < 0.01; *** p < 0.001;**** p < 0.0001

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Western Blot, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Apoptotic effects of CB2R ligands in GC cell lines. A ) Muse Annexin V Cell Assay for HGC27-S/R and AGS cell lines evaluated after 48 h of treatment with 1 µM or 6 µM of AM630 and 1 µM or 10 µM of CC48 , Fi9 , ASF151 and compound 1 . Results expressed as relative apoptosis rate compared to control cells and derived from three independent experiments were expressed as mean ± SD and plotted in the corresponding graphs. * p < 0.05; ** p < 0.01; B ) Representative western blotting analyses performed in HGC27-S/R and AGS cells regarding the expression of PPRγ, P-JUN/JUN, P-JNK/JNK and cleaved caspase 3/7. Actin was used as a normalizer of the protein extracts. C ) Statistical charts reporting the results from three independent western blotting experiments performed in HGC27-S/R and AGS cells and expressed as means ± SD, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Control, Derivative Assay, Western Blot, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Apoptotic profile of CB2R ligands and PTX in HGC27-S/R and NCI-N87 cells. The Muse caspase 3/7 activation Cell Assay was assessed after 48 h of both single drug treatments with 4 nM PTX, 6 µM AM630 , 10 µM CC48 , Fi9 , ASF151 and compound 1 , and after combined treatment of PTX with CB2R target compounds. A ) Representative flow cytometry charts of HGC27-S/R are shown. Four cell populations can be distinguished, namely the percentage of live cells (bottom left quadrant), cells in early apoptosis (bottom right quadrant), in late apoptosis (top right quadrant) and dead cells (top left quadrant). B ) The results derived from three independent experiments on HGC27-S, HGC27-R and NCI-N87 cell lines are expressed as means ± SD and reported in the relative graphs. Statistical analysis was conducted, comparing the values obtained using single drug treatment to those of corresponding untreated cells and the combined treatments to those of the single treatments, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Activation Assay, Flow Cytometry, Derivative Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CC48 a new CB2R agonist/FAAH inhibitor dual drug blocks gastric cancer progression and overcomes paclitaxel resistance

doi: 10.1186/s13046-025-03476-7

Figure Lengend Snippet: Effects of CB2R ligands on inhibition of cell migration and VEGFA secretion. A ) Scratch assay evaluated on HGC27-S/R and AGS treated with 1 and 6 µM AM630 and 1 and 10 µM CC48 , Fi9 , ASF151 and 1 . Cells were microscopically analyzed at the time of scratching (T0) and after 24 h (T1). The relative migration rate was calculated by placing the percentage migration of control cells at time T1 equal to 1 and comparing the percentage migration of cells after each drug treatment with this value. The experiments were performed in triples and the average SD values were plotted in the relative graph. * p < 0,05; ** p < 0,01; B ) Representative western blotting analyses performed in HGC27- S/R and AGS cells regarding the expression of P-βcatenin/βcatenin, vimentin and P-cofillin/cofillin. Actin was used as a normalization of the protein extracts; C ) Effects of CB2R ligands on VEGFA/VEGFC secretion. The ELISA assay was assessed on HGC27-S/R and AGS treated with 1 and 6 µM AM630 and 1 and 10 µM CC48 , Fi9 , ASF151 and 1 . The concentration of VEGFA was determined in the medium and normalized for the cell number. The values ± SD, obtained from three independent experiments expressed as pg/mL were shown in the relative graphs.** p < 0,01; *** p < 0,001

Article Snippet: The two compounds CC48 and Fi9 are classified as MultiTarget Directed Ligands (MTDLs) due to their dual activity.

Techniques: Inhibition, Migration, Wound Healing Assay, Control, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay