Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: TIMP1 expression in several types of cancer. (A) UALCAN database was used to analyze the expression levels of TIMP1 in several types of pan-cancer tissues (indicated by the red box) and their corresponding normal tissues (indicated by the blue box). (B) Expression of TIMP1 was significantly upregulated in colon cancer tissues compared with that in normal tissues. (C) Expression level of TIMP1 was significantly upregulated in colon tissues compared with that in normal tissues, based on TCGA data from the Gene Expression Profiling Interactive Analysis 2.0 database. (D) Expression level of TIMP1 was significantly upregulated in colon tissues (G2) compared with in normal tissues (G1) in the GSE24550 dataset. (E) Protein expression of TIMP1 in colon tissues from CPTAC data based on the UALCAN database. *P<0.05; ****P<0.0001. TIMP1, tissue inhibitor of matrix metalloproteinase 1; UALCAN, University of Alabama at Birmingham CANcer data analysis Portal; TCGA, The Cancer Genome Atlas; CPTAC, Clinical Proteomic Tumor Analysis Consortium; TPM, transcripts per million; G, grade; COAD, colon adenocarcinoma.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Expressing, Gene Expression

Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: Association between TIMP1 expression and survival prognosis of patients with CRC. (A) Association between TIMP1 expression and the overall survival of patients with CRC was analyzed using the GEPIA 2.0 database. (B) Association between TIMP1 expression and the overall survival of patients with colon cancer was analyzed using the University of Alabama at Birmingham CANcer data analysis Portal database. (C) Kaplan-Meier survival curve for TIMP1 in CRC-The Cancer Genome Atlas data, with different groups compared using the log-rank test. (D) Association between TIMP1 expression and the disease-free survival of patients with CRC was analyzed using the GEPIA 2.0 database. TIMP1, tissue inhibitor of matrix metalloproteinase 1; CRC, colorectal cancer; GEPIA, Gene Expression Profiling Interactive Analysis; HR, hazard ratio; CI, confidence interval; COAD, colon adenocarcinoma.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Expressing, Gene Expression

Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: TIMP1 serves as a prognostic factor for overall survival in CRC. (A) Univariate and (B) multivariate Cox analyses of TCGA data of patients with CRC for TIMP1 expression and clinical characteristics. (C) Nomogram formulated to predict 1-year overall survival for patients with CRC from TCGA data. (D) Calibration curve for the overall survival nomogram model in the discovery cohort, where the diagonal dashed line denotes the ideal nomogram, and the red line represent the observed 1-year nomograms. TIMP1, tissue inhibitor of matrix metalloproteinase 1; CRC, colorectal cancer; TCGA, The Cancer Genome Atlas; HR, hazard ratio; CI, confidence interval; p, pathological; T, tumor; N, node; M, metastasis.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Expressing

Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: Association between TIMP1 expression and immunity in CRC. Heatmap illustrates the correlation analysis of TIMP1 expression with (A) TIMER and (B) EPIC scores. Red signifies a positive correlation, whilst blue indicates a negative correlation. The intensity of the red or blue color reflects the strength of the correlation, and the size of the circle indicates the magnitude of the correlation. In the schematic, a red line denotes a negative correlation between model scores/gene expression and immune scores, whereas a green line denotes a positive correlation. (C) Spearman's correlation analysis of TIMP1 expression with CD4 + and CD8 + T cells in CRC from The Cancer Genome Atlas database. (D) Spearman's correlation analysis of TIMP1 expression with TMB/MSI score distribution is shown. TIMP1, tissue inhibitor of matrix metalloproteinase 1; CRC, colorectal cancer; TMB, tumor mutation burden; MSI,/microsatellite instability; NK, natural killer.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Expressing, Gene Expression, Mutagenesis

Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: Correlation of TIMP1 and signaling pathways in CRC. Spearman's correlation analysis plots reveal the correlation between pathway scores and TIMP1 expression in CRC samples from The Cancer Genome Atlas database. TIMP1, tissue inhibitor of matrix metalloproteinase 1; CRC, colorectal cancer; TPM, transcripts per million; EMT, epithelial-mesenchymal transition; ECM, extracellular matrix.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Protein-Protein interactions, Expressing

Journal: Oncology Letters

Article Title: TIMP1 expression in colorectal cancer: Linking prognosis, tumor immunity and molecular pathways

doi: 10.3892/ol.2025.15390

Figure Lengend Snippet: TIMP1 protein expression in the colorectal cancer cohort in the present study. (A) Representative images of IHC analysis of TIMP1 protein expression of NAT (n=78) and CC (n=78) tissue sections. Scale bar, 100 µm. (B) TIMP1 protein expression based on their staining index in NAT and CC specimens. (C) Patients with CC with high TIMP1 expression (IHC score ≥4) had a significantly worse prognosis than patients with low TIMP1 expression (IHC score <4). ***P<0.001. TIMP1, tissue inhibitor of matrix metalloproteinase 1; IHC, immunohistochemistry; NAT, nonmalignant adjacent tissues; CC, colon cancer.

Article Snippet: The sections were then incubated with anti-TIMP1 antibodies (1:300 dilution; cat. no. 16644-1-AP; Proteintech Group, Inc.) overnight at 4°C, succeeded by incubation with a biotinylated goat anti-rabbit secondary antibody working solution (1:500 dilution; cat. no. SA1020; Boster Biological Technology Co. Ltd.) at 37°C for 30 min. Immunodetection was subsequently executed employing the Dako EnVision detection system (Agilent Technologies, Inc.).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Journal of Inflammation Research

Article Title: Tellimagrandin II Stimulates Inflammasomes by Causing an Accumulation of 3-Aminopropanal, Which Promotes Apoptosis of Endometriotic Cells While Inhibiting Invasion

doi: 10.2147/JIR.S558146

Figure Lengend Snippet: Western blot analysis of EECs treated with Tellimagrandin II: ( a ) protein immunoblot; ( b ) ALDH7A1; ( c ) SMOX; ( d ) NLRP3; ( e ) Caspase-1; ( f ) β-catenin; ( g ) cleaved Caspase-3; ( h ) BAX; ( i ) MMP2; ( j ) TIMP1. Each experiment was repeated three times. *:P <0.05; **:P <0.01; ***:P <0.001; ****P <0.0001.

Article Snippet: Tellimagrandin II (Yongjian: B02179 ), Spermine Oxidase (SMOX) Polyclonal antibody (proteintech: 15052-1-AP), Aldehyde Dehydrogenase 7A1 (ALDH7A1) Polyclonal antibody (proteintech: 10368-1-AP), Anti-NLR Family, Pyrin Domain Containing Protein 3 (NLRP3) Rabbit pAb (Servicebio: GB114320-50), Recombinant Anti-beta Catenin (β-Catenin) antibody (Servicebio: GB150016-100), Anti-Matrix Metallopeptidase 2 (MMP2) Rabbit pAb (Servicebio: GB11130-100), Anti- Tissue Inhibitors of Metalloproteinase 1 (TIMP1) Rabbit pAb (proteintech: 16644-1-AP), Anti-Caspase-3 Rabbit pAb (Servicebio: GB115600-100), Anti-Bcl-2-associated X(Bax) Rabbit pAb (Servicebio: GB11690-100), Anti-Caspase-1 Rabbit pAb (Servicebio: GB11383-100), DMEM/F12 (Glutamine and HEPES)(M&C GENE: CM10092), Fetal Bovine Serum Gold (YEASEN: 40130ES76), penicillin streptomycin mixture (Servicebio: P1400), 1×PBS buffer (Solarbio: P1020), trypsin-EDTA digestion solution (Phenol Red)(Solarbio: T1320), CCK-8 kit (Sola rbio: CA1210), Annexin V-FITC Apoptosis detection kit (Beyotime: C1062S), 96-well cell culture plate (Servicebio:) CCP-96H, SWE matrix adhesive (Solarbio: G4131-5ML), crystal violet staining solution (Solarbio: G1014-50ML), paraformaldehyde fixative (Solarbio: G1101-500ML), Transwell chamber (Corning: WG3422) Cell culture flasks (Corning: 430639), 12Z EECs line (Bioharbor: LH-H337), DMSO (Servicebio: GC203006-10mL), Propidium Iodide (proteintech: CM18819), Flow cytometer (BD: FACSAria II), Gel imaging system (Alpha: 2200) Electrophoresis apparatus (Thermo: EC250-90), Thermo Corporation, USA; Pendulum type decolorizing shaker (model: DS-2S100), Nanodrop micro spectrophotometer (Illumina: Nanodrop2000), sequencing platform machine (Illumina: NovaSeq X Plus).

Techniques: Western Blot

Journal: Chinese Medicine

Article Title: Colla Corii Asini regulate collagen regeneration in UV exposure-induced skin photoaging in mice

doi: 10.1186/s13020-025-01175-1

Figure Lengend Snippet: CCA regulates the dynamic balance of collagen synthesis and degradation via the TGFβ-SMAD/MMPs pathway. A , B RT-qPCR analysis and statistical evaluation of gene expression in the TGFβ-SMAD/MMPs pathway ( A ). RT-qPCR results showing mRNA expression levels of TGFBR2, TGFBR1, SMAD2, SMAD3, SMAD4, MMP1, and MMP10 in mouse skin after 4 weeks of treatment with control, model, CCA, or VC (n = 3) ( B ). RT-qPCR results showing mRNA expression levels of TGFBR2, TGFBR1, SMAD2, SMAD3, SMAD4, MMP1, and MMP10 in mouse skin after 9 weeks of treatment with control, model, CCA, or VC (n = 4). C, D WB analysis and statistical evaluation of protein expression in the TGFβ-SMAD/MMPs pathway ( C ). WB results showing protein expression levels of TGFBR2, TGFBR1, SMAD2/3, SMAD4, MMP1, and TIMP1 in mouse skin after 4 weeks of treatment with control, model, CCA, or VC, with GAPDH used as a loading control (n = 3) ( D ). WB results showing protein expression levels of TGFBR2, TGFBR1, SMAD2/3, SMAD4, MMP1, and TIMP1 in mouse skin after 9 weeks of treatment with control, model, CCA, or VC, with GAPDH used as a loading control (n = 3).Data are presented as mean ± SEM (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Specific proteins were detected using the following primary antibodies: Collagen I (14695-1-AP, Proteintech Group Inc., 1:1000), Collagen III (22734-1-AP, Proteintech Group Inc., 1:1000), TGFBR1 (30117-1-AP, Proteintech Group Inc., 1:1000), TGFBR2 (66636-1-Ig, Proteintech Group Inc., 1:1000), SMAD2 (12570-1-AP, Proteintech Group Inc., 1:1000), SMAD3 (66599-1-Ig, Proteintech Group Inc., 1:1000), SMAD2/3 (AP0548, ABclonal Biotechnology Co., Ltd., China, 1:1000), SMAD4 (10231-1-AP, Proteintech Group Inc., 1:1000), MMP1 (10371-2-AP, Proteintech Group Inc., 1:1000), TIMP1 (16644-1-AP, Proteintech Group Inc., 1:1000), MAPK14 (14064-1-AP, Proteintech Group Inc., 1:1000), JUN (24909-1-AP, Proteintech Group Inc., 1:1000), FOS (66590-1-Ig, Proteintech Group Inc., 1:1000), P16 (10883-1-AP, Proteintech Group Inc., 1:1000), P21 (10355-1-AP, Proteintech Group Inc., 1:1000), and P53 (10442-1-AP, Proteintech Group Inc., 1:1000), GAPDH (10494-1-AP, Proteintech Group Inc., 1: 1000),DYKDDDDK tag (66008-4-Ig, Proteintech Group Inc., 1:5000).

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Control

Journal: Chinese Medicine

Article Title: Colla Corii Asini regulate collagen regeneration in UV exposure-induced skin photoaging in mice

doi: 10.1186/s13020-025-01175-1

Figure Lengend Snippet: Pharmacological effects and colocalization analysis of CCA peptides. A Schematic diagram of peptide experiment (The Figure was generated using Figuredraw). B PPI analysis of CCA peptides with the TGFβ-SMAD pathway and visualization of AlphaFold3-predicted structures. Left: PPI analysis of CCA peptides with the TGFβ-SMAD pathway. Right: AlphaFold3-predicted structures of YYTSASGDEMVSLK and HFSVEGQLEFR. C, D Pharmacological validation of peptides ( C ). RT-qPCR analysis of Collagen I mRNA expression levels after treatment with different concentrations of YYTSASGDEMVSLK and HFSVEGQLEFR (n = 6) ( D ). WB analysis showing protein expression levels of P21, P16, Collagen I, and Collagen III in different groups, with GAPDH used as the loading control (n = 3). E, F Regulation of TGFBR1 and TGFBR2 gene and protein expression by YYTSASGDEMVSLK to activate the TGFβ-SMAD pathway ( E ). RT-qPCR analysis of TGFBR1 and TGFBR2 mRNA expression levels in photoaged BJ cells treated with 0.25 mg/mL YYTSASGDEMVSLK (n = 6) ( F ). WB analysis showing protein expression levels of TGFBR2, TGFBR1, SMAD2/3, pSMAD2/3, SMAD4, MMP1, and TIMP1 in different groups, with GAPDH used as the loading control (n = 3). G Visualization of AlphaFold3-predicted interactions between YYTSASGDEMVSLK and TGFBR1/TGFBR2. orange: AlphaFold3-predicted PLDDT values and 3D structures of YYTSASGDEMVSLK bound to TGFBR1 (PDB ID: 4xm0) and TGFBR2 (PDB ID: 5qin). green: schematic representation of colocalization of YYTSASGDEMVSLK with TGFBR1 and TGFBR2 on the cell membrane. H, I Colocalization analysis of YYTSASGDEMVSLK with TGFBR1 and TGFBR2 ( H ). WB analysis showing binding of YYTSASGDEMVSLK-Flag to TGFBR1 and TGFBR2 in BJ cells incubated with 0.25 mg/mL YYTSASGDEMVSLK-Flag for 24 h ( I ). STEDYCON imaging showing colocalization of YYTSASGDEMVSLK-FITC (green) with TGFBR1 and TGFBR2 (red). Yellow indicates colocalization, and nuclei are stained with DAPI (blue) (scale bar = 20 µm). Data are presented s mean ± SEM (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Specific proteins were detected using the following primary antibodies: Collagen I (14695-1-AP, Proteintech Group Inc., 1:1000), Collagen III (22734-1-AP, Proteintech Group Inc., 1:1000), TGFBR1 (30117-1-AP, Proteintech Group Inc., 1:1000), TGFBR2 (66636-1-Ig, Proteintech Group Inc., 1:1000), SMAD2 (12570-1-AP, Proteintech Group Inc., 1:1000), SMAD3 (66599-1-Ig, Proteintech Group Inc., 1:1000), SMAD2/3 (AP0548, ABclonal Biotechnology Co., Ltd., China, 1:1000), SMAD4 (10231-1-AP, Proteintech Group Inc., 1:1000), MMP1 (10371-2-AP, Proteintech Group Inc., 1:1000), TIMP1 (16644-1-AP, Proteintech Group Inc., 1:1000), MAPK14 (14064-1-AP, Proteintech Group Inc., 1:1000), JUN (24909-1-AP, Proteintech Group Inc., 1:1000), FOS (66590-1-Ig, Proteintech Group Inc., 1:1000), P16 (10883-1-AP, Proteintech Group Inc., 1:1000), P21 (10355-1-AP, Proteintech Group Inc., 1:1000), and P53 (10442-1-AP, Proteintech Group Inc., 1:1000), GAPDH (10494-1-AP, Proteintech Group Inc., 1: 1000),DYKDDDDK tag (66008-4-Ig, Proteintech Group Inc., 1:5000).

Techniques: Generated, Biomarker Discovery, Quantitative RT-PCR, Expressing, Control, Membrane, Binding Assay, Incubation, Imaging, Staining