Journal: bioRxiv

Article Title: CASTOR1 : A Novel Tumor Suppressor Linking mTORC1 and KRAS Pathways in Tumorigenesis and Resistance to KRAS-Targeted Therapies in Non-Small Cell Lung Cancer

doi: 10.1101/2025.04.23.650349

Figure Lengend Snippet: (A) Representative H&E images of lungs collected from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice 5 months after intranasal inoculation with Adeno-Cre (left panel). Tumor incidence, size and burden (tumor area/total lung area ×100) were quantified (left, middle and right panels, respectively). Data presented as mean +/- standard error of the mean (SEM). Statistical analysis was performed by nested-Anova and Student’s t-test. (B) Representative IHC staining of CASTOR1 in lung tumors from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice. (C) Representative IHC staining images of Ki67 in lung tumors from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice (left panel) and quantification of Ki67-positive cells (right panel). Statistical analysis was performed by nested-Anova. (D and E) Representative IHC staining images of p-4EBP1 (D) and p-S6 (E) of lung tumors from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice (left panels) and quantification of positive cells (right panels). Statistical analysis was performed by nested-Anova. (F) Immunoblotting analysis of mTORC1 downstream targets S6K and 4EBP1, KRAS downstream target ERK and AKT, and their phosphorylated forms in normal lung tissues from KRAS WT ;C1 WT and KRAS WT ;C1 KO mice, and tumors from KRAS G12D ;C1 WT and KRAS G12D ;C1 KO mice. Three samples per group were analyzed. (G). Representative IHC staining images of p-ERK in lung tumors from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice (left panel) and quantification of positive cells (right panel). Statistical analysis was performed by Student’s t-test.

Article Snippet: Primary antibodies include CASTOR1 (Abclonal, cat. no. A2071C, 1:50 dilution), p-ERK (Cell Signaling Technology, cat. no.4370, 1:100 dilution), p-4EBP1 (Cell Signaling Technology, cat. no.2855, 1:100), p-S6K (Abclonal, cat. no.AP0502, 1:100 dilution), p-S6 (Cell Signaling Technology, cat. no.2211, 1:100), and pCASTOR1 (1:100).

Techniques: Immunohistochemistry, Western Blot

Journal: bioRxiv

Article Title: CASTOR1 : A Novel Tumor Suppressor Linking mTORC1 and KRAS Pathways in Tumorigenesis and Resistance to KRAS-Targeted Therapies in Non-Small Cell Lung Cancer

doi: 10.1101/2025.04.23.650349

Figure Lengend Snippet: (A-C) CASTOR1 ablation enhances resistance to KRAS G12D -specific inhibitor MRTX1133 in tumors. Mice were intranasally inoculated with Adeno-Cre for 4 weeks, then treated with MRTX1133 of vehicle for 4 weeks. Representative H&E images of lungs collected from KRAS G12D ;C1 WT or KRAS G12D ;C1 KO mice (A). Tumor size (B) and burden (tumor area/total lung area ×100) (C) were quantified. Data presented as mean +/- standard error of the mean (SEM). Statistical analysis was performed by nested-Anova and Student’s t-test. (D) KRAS G12D ;C1 WT tumor organoids had heterogeneous response to MRTX1133. Representative pictures of nine KRAS G12D ;C1 WT organoids after one week of treatment with MRTX1133 or vehicle. (E) Immunoblotting analysis of KRAS G12D protein, mTORC1 downstream targets S6K and 4EBP1, and KRAS downstream target ERK in KRAS G12D ;C1 WT tumor organoids. (F) KRAS G12D ;C1 KO tumor organoids were resistant to MRTX1133. Representative pictures of four KRAS G12D ;C1 KO organoids after one week of treatment with MRTX1133 or vehicle. (G) Comparison of sensitivity to MRTX1133 between organoids derived from KRAS G12D ;C1 WT and KRAS G12D ;C1 KO tumors. (H-I) mTORC1 inhibitor Rapamycin and PI3K inhibitor BGT226 sensitized KRAS G12D ;C1 WT and KRAS G12D ;C1 KO organoids to MRTX1133. Representative pictures of organoids treated with mono- or combined therapy of MRTX1133 and Rapamycin (H) or BGT226 (I).

Article Snippet: Primary antibodies include CASTOR1 (Abclonal, cat. no. A2071C, 1:50 dilution), p-ERK (Cell Signaling Technology, cat. no.4370, 1:100 dilution), p-4EBP1 (Cell Signaling Technology, cat. no.2855, 1:100), p-S6K (Abclonal, cat. no.AP0502, 1:100 dilution), p-S6 (Cell Signaling Technology, cat. no.2211, 1:100), and pCASTOR1 (1:100).

Techniques: Western Blot, Comparison, Derivative Assay

Journal: Intervirology

Article Title: Molecular Biological Mechanisms of Action of Chrysophanol in Hepatic Stellate Cells Activated by Hepatic B Virus X Based on Network Pharmacology

doi: 10.1159/000542355

Figure Lengend Snippet: Cho targets HBx to inhibit the upregulation of mTOR, p-mTOR, S6, p-S6, PPAR-α, p-PPAR-α, and CYP27 expressions and activation of liver stellate T6 cells. Left panel indicated the changes in the expression of hepatitis B virus X protein (HBx) (FLAG), mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), S6, phosphorylated S6 (p-S6), peroxisome proliferator-activated receptor (PPAR)-α, phosphorylated-PPAR-α (p-PPAR-α), CYP27, α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), and Integrin-β1. β-actin was used as an internal control. Right panel indicated the quantitative images demonstrating the levels of specific proteins determined using ImageJ software. Error bars display standard deviations from three independent replications ( n = 3). * p < 0.05; ** p < 0.01. Cho = 30 μm Cho treatment.

Article Snippet: The following antibodies were used: rabbit mammalian target of rapamycin (mTOR) antibody (1:1,000, #A2445; ABclonal, Woburn, MA, USA), rabbit phospho-mTOR antibody (1:1,000, #AP0094; ABclonal), rabbit S6 Kinase 1 antibody (1:1,000; #A2190; ABclonal), rabbit phospho-S6 Kinase 1 antibody (1:1,000; #AP1105; ABclonal), rabbit peroxisome proliferator-activated receptor (PPAR)-α (1:1,000, #A18252; ABclonal), rabbit phospho-PPAR-α (1:1,000, #AF8054; Affinity Biosciences, OH, USA), rabbit CYP27 (1:1,000, # A23250; ABclonal), rabbit CTGF (1:1,000, #A11456; ABclonal), rabbit Integrin-β1 (1:1,000, #A19072; ABclonal), rabbit FLAG (1:1,000, #AE004; ABclonal) for HBx, rabbit β-actin (1:5,000, #AC038; ABclonal), mouse α-SMA (1:1,000, #14968; Cell Signaling Technology, Danvers, MA, USA), horseradish peroxidase-conjugated goat anti-rabbit IgG (1:7,500; Cell Signaling Technology), and horseradish peroxidase-conjugated goat anti-mouse IgG (1:7,500; Cell Signaling Technology).

Techniques: Activation Assay, Expressing, Virus, Control, Software

Journal: Intervirology

Article Title: Molecular Biological Mechanisms of Action of Chrysophanol in Hepatic Stellate Cells Activated by Hepatic B Virus X Based on Network Pharmacology

doi: 10.1159/000542355

Figure Lengend Snippet: Cho inhibits the upregulation of mTOR, p-mTOR, S6, p-S6, PPAR-α, p-PPAR-α, and CYP27 expressions activation of liver stellate cells in TAA-induced liver fibrosis mouse model and HSC-LX2 cells. Upper left panel indicated the changes in the expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), S6, phosphorylated S6 (p-S6), peroxisome proliferator-activated receptor (PPAR)-α, phosphorylated-PPAR-α (p-PPAR-α), CYP27, α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), and Integrin-β1 in TAA-induced liver fibrosis mouse model. β-actin was used as an internal control. Upper right panel indicated the changes in the expression of hepatitis B virus X protein (HBx) (FLAG), mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), S6, phosphorylated S6 (p-S6), peroxisome proliferator-activated receptor (PPAR)-α, phosphorylated-PPAR-α (p-PPAR-α), CYP27, α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), and Integrin-β1 in HSC-LX2 cells.. β-actin was used as an internal control. Lower pannel indicated the quantitative images demonstrating the levels of specific proteins determined using ImageJ software. Error bars display standard deviations from three independent replications ( n = 3). * p < 0.05. Cho = 30 μm Cho treatment.

Article Snippet: The following antibodies were used: rabbit mammalian target of rapamycin (mTOR) antibody (1:1,000, #A2445; ABclonal, Woburn, MA, USA), rabbit phospho-mTOR antibody (1:1,000, #AP0094; ABclonal), rabbit S6 Kinase 1 antibody (1:1,000; #A2190; ABclonal), rabbit phospho-S6 Kinase 1 antibody (1:1,000; #AP1105; ABclonal), rabbit peroxisome proliferator-activated receptor (PPAR)-α (1:1,000, #A18252; ABclonal), rabbit phospho-PPAR-α (1:1,000, #AF8054; Affinity Biosciences, OH, USA), rabbit CYP27 (1:1,000, # A23250; ABclonal), rabbit CTGF (1:1,000, #A11456; ABclonal), rabbit Integrin-β1 (1:1,000, #A19072; ABclonal), rabbit FLAG (1:1,000, #AE004; ABclonal) for HBx, rabbit β-actin (1:5,000, #AC038; ABclonal), mouse α-SMA (1:1,000, #14968; Cell Signaling Technology, Danvers, MA, USA), horseradish peroxidase-conjugated goat anti-rabbit IgG (1:7,500; Cell Signaling Technology), and horseradish peroxidase-conjugated goat anti-mouse IgG (1:7,500; Cell Signaling Technology).

Techniques: Activation Assay, Expressing, Control, Virus, Software

Journal: Intervirology

Article Title: Molecular Biological Mechanisms of Action of Chrysophanol in Hepatic Stellate Cells Activated by Hepatic B Virus X Based on Network Pharmacology

doi: 10.1159/000542355

Figure Lengend Snippet: Treatment with mTOR activator MHY-1485 and PPAR-α agonist WY-14643 counteracted the effects of Cho on HBx-activated HSCs. Upper panel indicates the changes in the expression of phosphorylated mammalian target of rapamycin (p-mTOR), α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), and Integrin-β1. β-actin was used as an internal control. Cho = 30 μm Cho treatment. MHY-1485 = 10 µm. Lower panel indicates the changes in the expression of phosphorylated peroxisome proliferator-activated receptor (p-PPAR-α), α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), and Integrin-β1. β-actin was used as an internal control. Cho = 30 μm Cho treatment. WY-14643 = 10 μm. Quantitative images demonstrating the levels of specific proteins determined using ImageJ software. Error bars display standard deviations from three independent replications ( n = 3). * p < 0.05.

Article Snippet: The following antibodies were used: rabbit mammalian target of rapamycin (mTOR) antibody (1:1,000, #A2445; ABclonal, Woburn, MA, USA), rabbit phospho-mTOR antibody (1:1,000, #AP0094; ABclonal), rabbit S6 Kinase 1 antibody (1:1,000; #A2190; ABclonal), rabbit phospho-S6 Kinase 1 antibody (1:1,000; #AP1105; ABclonal), rabbit peroxisome proliferator-activated receptor (PPAR)-α (1:1,000, #A18252; ABclonal), rabbit phospho-PPAR-α (1:1,000, #AF8054; Affinity Biosciences, OH, USA), rabbit CYP27 (1:1,000, # A23250; ABclonal), rabbit CTGF (1:1,000, #A11456; ABclonal), rabbit Integrin-β1 (1:1,000, #A19072; ABclonal), rabbit FLAG (1:1,000, #AE004; ABclonal) for HBx, rabbit β-actin (1:5,000, #AC038; ABclonal), mouse α-SMA (1:1,000, #14968; Cell Signaling Technology, Danvers, MA, USA), horseradish peroxidase-conjugated goat anti-rabbit IgG (1:7,500; Cell Signaling Technology), and horseradish peroxidase-conjugated goat anti-mouse IgG (1:7,500; Cell Signaling Technology).

Techniques: Expressing, Control, Software