Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: This schematic diagram provides an overview of the study’s integrated analytical framework. The workflow begins with data acquisition from public databases, followed by the integration of single-cell RNA sequencing and bulk multi-omics datasets. Metastasis-associated fibroblast subpopulations are then identified through clustering and functional annotation. Key gene modules are extracted using hdWGCNA, and a prognostic risk model is constructed and validated across independent cohorts. Finally, core genes—such as TPP1 —are experimentally validated using in vitro assays to confirm their functional relevance in gastric cancer progression.

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Single Cell, RNA Sequencing, Biomarker Discovery, Functional Assay, Construct, In Vitro

Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: Abnormal expression of TPP1 was verified. ( A ) Boruta algorithm identified six key prognostic signature genes, with TPP1 ranking as the top contributor. ( B ) Pan-cancer differential expression analysis revealed significant overexpression of TPP1 across multiple malignancies. ( C , D ) Bar plots show that TPP1 expression is significantly elevated in gastric cancer tissues based on both unpaired and paired comparisons. ( E ) Kaplan–Meier survival analysis demonstrated that high TPP1 expression correlates with poorer overall survival in gastric cancer patients. ( F ) Immunohistochemical staining confirmed upregulated TPP1 protein expression in gastric cancer tissues (HPA: The Human Protein Atlas). ( G ) Western blot analysis further validated increased TPP1 protein levels in tumor samples. ( H ) qRT-PCR analysis confirmed that TPP1 mRNA expression was significantly elevated in gastric cancer tissues (n = 4). Data are presented as mean ± standard deviation (SD). Statistical comparisons were performed using Student’s t -test unless otherwise indicated. Significance levels were defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Expressing, Quantitative Proteomics, Over Expression, Immunohistochemical staining, Staining, Western Blot, Quantitative RT-PCR, Standard Deviation

Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: Silencing of TPP1 attenuates tumorigenic properties and enhances apoptosis in gastric cancer cells. ( A ) qRT-PCR analysis confirmed elevated TPP1 expression in gastric cancer cell lines (n = 3). ( B ) Western blot analysis validated high TPP1 protein levels across gastric cancer cell lines (n = 3). ( C ) qRT-PCR confirmed effective knockdown of TPP1 following siRNA transfection (n = 3). ( D ) Colony formation assays demonstrated a significant reduction in clonogenic capacity upon TPP1 silencing (n = 3). ( E ) Flow cytometry analysis using Annexin V/DAPI staining revealed increased apoptosis in TPP1-depleted cells (n = 3). ( F ) Transwell invasion assays showed a marked decrease in invasive capacity following TPP1 knockdown (n = 3). ( G ) CCK-8 assays indicated impaired cell viability in TPP1-silenced gastric cancer cells (n = 3). Data are presented as mean ± standard deviation (SD), and all experiments were performed in triplicate. Statistical significance was assessed using Student’s t-test unless otherwise specified: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Knockdown, Transfection, Flow Cytometry, Staining, CCK-8 Assay, Standard Deviation

Journal: Cell metabolism

Article Title: FBW7 Mediates Senescence and Pulmonary Fibrosis through Telomere Uncapping.

doi: 10.1016/j.cmet.2020.10.004

Figure Lengend Snippet: Figure 2. FBW7 Mediates TPP1 Multisite Polyubiquitination and Turnover via GSK3b Signaling in Stress Response (A) Effect of various E3 ubiquitin ligase gene KDs on TPP1 in A549 cells exposed to H2O2, BLM, or IR by IB. Cells transfected for 48 h were exposed to BLM (200 mM, 16 h), H2O2 (1 mM), or IR (8 Gy).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 Thermo Cat#A27034 Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 555 Thermo Cat#A27039 Anti-rabbit IgG-Peroxidase antibody produced in goat Sigma Cat#A6154 Anti-Rap1 antibody IMGENEX Cat#IMG-289 Anti-SP-C antibody (for western blot) Abcam Cat#ab90716 Anti-SP-C antibody (for immunofluorescence) Santa Cruz Biotechnology Cat#sc-7706 Anti-T1a antibody Biolegend Cat#127408 Anti-Ter119 antibody Biolegend Cat#116204 Anti-TIN2 antibody GeneTex Cat59B388 Anti-TPP1 antibody (for immunofluorescence) Novus Cat#NBP2-45475 Anti-TPP1 antibody (for western blot) Abcam Cat#ab112050 and ab54685 Anti-TRF1 antibody Cell Signaling Technology Cat#3529 Anti-TRF2 antibody Novus Cat#NB100-56506 Anti-a-SMA antibody Sigma Cat#A2547 Anti-a-Tubulin antibody Cell Signaling Technology Cat#2144 Peroxidase-AffiniPure goat anti mouse IgG Jackson Cat#115-035-062 Bacterial and Virus Strains Cre lentivirus Genomeditech Cat#GM-0220CR02 E. coli (DH5a) TransGen Biotech Cat#CD201-01 pEF-1a-TPP1 lentivirus GenePharma N/A pLMCS-FBW7a lentivirus Invitrogen (Shanghai) N/A pLMCS-FBW7g lentivirus Invitrogen (Shanghai) N/A Chemicals, Peptides, and Recombinant Proteins Bleomycin Sigma Cat#5507 BrdU Sigma Cat#B5002 CHIR99021 Sigma Cat#SML1046 Colcemid Thermo Cat# 15212012 Cycloheximide Sigma Cat#239763 Digoxin Roche Cat#11585614910 Dispase II Sigma Cat#4942078001 DMSO Sigma Cat#D8418 Dynabeads MyOne streptavidin T1 magnetic beads Thermo Cat#65601 Flag-coupled magnetic beads Sigma Cat#M8823 Glutathione-Sepharose 4B Sigma Cat#GE17-0756-01 H2O2 (3%) Sigma Cat#88597 Lipofectamine 2000 Thermo Cat#11668019 Masson Trichrome Sigma Cat#HT153 MG132 Sigma Cat#M8699 Peptides KE BIOCHEM See Table S5 PNA probe Panagene Cat#F1001,F1002 Streptavidin-PE Biolegend Cat#405204 TC-G24 TOCRIS Cat#4353 (Continued on next page) e2 Cell Metabolism 32, 1–18.e1–e9, November 3, 2020

Techniques: Ubiquitin Proteomics, Transfection

Journal: Cell metabolism

Article Title: FBW7 Mediates Senescence and Pulmonary Fibrosis through Telomere Uncapping.

doi: 10.1016/j.cmet.2020.10.004

Figure Lengend Snippet: Figure 4. FBW7 Regulates Telomere Uncapping and Cell Proliferation in Response to H2O2 or IR in a TPP1-Dependent Manner (A) Effect of FBW7 KD on TIFs determined by IF-FISH in HeLa cells transfected with the indicated siRNAs for 48 h. (B and C) Effect of FBW7 KD on telomere length by flow-FISH in HT1080 cells (B) and on telomere numbers by metaphase FISH per chromosome in HeLa cells (C).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 Thermo Cat#A27034 Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 555 Thermo Cat#A27039 Anti-rabbit IgG-Peroxidase antibody produced in goat Sigma Cat#A6154 Anti-Rap1 antibody IMGENEX Cat#IMG-289 Anti-SP-C antibody (for western blot) Abcam Cat#ab90716 Anti-SP-C antibody (for immunofluorescence) Santa Cruz Biotechnology Cat#sc-7706 Anti-T1a antibody Biolegend Cat#127408 Anti-Ter119 antibody Biolegend Cat#116204 Anti-TIN2 antibody GeneTex Cat59B388 Anti-TPP1 antibody (for immunofluorescence) Novus Cat#NBP2-45475 Anti-TPP1 antibody (for western blot) Abcam Cat#ab112050 and ab54685 Anti-TRF1 antibody Cell Signaling Technology Cat#3529 Anti-TRF2 antibody Novus Cat#NB100-56506 Anti-a-SMA antibody Sigma Cat#A2547 Anti-a-Tubulin antibody Cell Signaling Technology Cat#2144 Peroxidase-AffiniPure goat anti mouse IgG Jackson Cat#115-035-062 Bacterial and Virus Strains Cre lentivirus Genomeditech Cat#GM-0220CR02 E. coli (DH5a) TransGen Biotech Cat#CD201-01 pEF-1a-TPP1 lentivirus GenePharma N/A pLMCS-FBW7a lentivirus Invitrogen (Shanghai) N/A pLMCS-FBW7g lentivirus Invitrogen (Shanghai) N/A Chemicals, Peptides, and Recombinant Proteins Bleomycin Sigma Cat#5507 BrdU Sigma Cat#B5002 CHIR99021 Sigma Cat#SML1046 Colcemid Thermo Cat# 15212012 Cycloheximide Sigma Cat#239763 Digoxin Roche Cat#11585614910 Dispase II Sigma Cat#4942078001 DMSO Sigma Cat#D8418 Dynabeads MyOne streptavidin T1 magnetic beads Thermo Cat#65601 Flag-coupled magnetic beads Sigma Cat#M8823 Glutathione-Sepharose 4B Sigma Cat#GE17-0756-01 H2O2 (3%) Sigma Cat#88597 Lipofectamine 2000 Thermo Cat#11668019 Masson Trichrome Sigma Cat#HT153 MG132 Sigma Cat#M8699 Peptides KE BIOCHEM See Table S5 PNA probe Panagene Cat#F1001,F1002 Streptavidin-PE Biolegend Cat#405204 TC-G24 TOCRIS Cat#4353 (Continued on next page) e2 Cell Metabolism 32, 1–18.e1–e9, November 3, 2020

Techniques: Transfection

Journal: Cell metabolism

Article Title: FBW7 Mediates Senescence and Pulmonary Fibrosis through Telomere Uncapping.

doi: 10.1016/j.cmet.2020.10.004

Figure Lengend Snippet: Figure 5. TPP1 KD Causes Telomere Shortening, AEC2 Stem Cell Senescence, and Pulmonary Fibrosis in Mice (A–G) TPP1 or FBW7 KD by weekly intratracheal shRNA lentiviruses for 4 times, before IB (A), FACS (B and C), examinations of lung function (D–F), and fibrotic marker HYP content (G). (H–J) Masson’s trichrome staining (H), telomere Q-FISH of 300–500 AEC2 cells (I), and IF for SPC-positive AEC2 cells (J) and HP1g-positive AEC2 cells (insets in J) in lung sections from 3 animals per group (20 fields per lung). *p < 0.05 versus control shRNA. (K) qPCR for FBW7, SPC, p21, a-SMA, and Col1a1 gene expression in the lung with TPP1 or FBW7 deficiency.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 Thermo Cat#A27034 Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 555 Thermo Cat#A27039 Anti-rabbit IgG-Peroxidase antibody produced in goat Sigma Cat#A6154 Anti-Rap1 antibody IMGENEX Cat#IMG-289 Anti-SP-C antibody (for western blot) Abcam Cat#ab90716 Anti-SP-C antibody (for immunofluorescence) Santa Cruz Biotechnology Cat#sc-7706 Anti-T1a antibody Biolegend Cat#127408 Anti-Ter119 antibody Biolegend Cat#116204 Anti-TIN2 antibody GeneTex Cat59B388 Anti-TPP1 antibody (for immunofluorescence) Novus Cat#NBP2-45475 Anti-TPP1 antibody (for western blot) Abcam Cat#ab112050 and ab54685 Anti-TRF1 antibody Cell Signaling Technology Cat#3529 Anti-TRF2 antibody Novus Cat#NB100-56506 Anti-a-SMA antibody Sigma Cat#A2547 Anti-a-Tubulin antibody Cell Signaling Technology Cat#2144 Peroxidase-AffiniPure goat anti mouse IgG Jackson Cat#115-035-062 Bacterial and Virus Strains Cre lentivirus Genomeditech Cat#GM-0220CR02 E. coli (DH5a) TransGen Biotech Cat#CD201-01 pEF-1a-TPP1 lentivirus GenePharma N/A pLMCS-FBW7a lentivirus Invitrogen (Shanghai) N/A pLMCS-FBW7g lentivirus Invitrogen (Shanghai) N/A Chemicals, Peptides, and Recombinant Proteins Bleomycin Sigma Cat#5507 BrdU Sigma Cat#B5002 CHIR99021 Sigma Cat#SML1046 Colcemid Thermo Cat# 15212012 Cycloheximide Sigma Cat#239763 Digoxin Roche Cat#11585614910 Dispase II Sigma Cat#4942078001 DMSO Sigma Cat#D8418 Dynabeads MyOne streptavidin T1 magnetic beads Thermo Cat#65601 Flag-coupled magnetic beads Sigma Cat#M8823 Glutathione-Sepharose 4B Sigma Cat#GE17-0756-01 H2O2 (3%) Sigma Cat#88597 Lipofectamine 2000 Thermo Cat#11668019 Masson Trichrome Sigma Cat#HT153 MG132 Sigma Cat#M8699 Peptides KE BIOCHEM See Table S5 PNA probe Panagene Cat#F1001,F1002 Streptavidin-PE Biolegend Cat#405204 TC-G24 TOCRIS Cat#4353 (Continued on next page) e2 Cell Metabolism 32, 1–18.e1–e9, November 3, 2020

Techniques: shRNA, Marker, Staining, Control, Gene Expression

Journal: Cell metabolism

Article Title: FBW7 Mediates Senescence and Pulmonary Fibrosis through Telomere Uncapping.

doi: 10.1016/j.cmet.2020.10.004

Figure Lengend Snippet: Figure 6. Peptidomimetic TELODIN Inhibits FBW7 Binding to TPP1 and Telomere Uncapping in Human Lung Epithelial Cells (A–C) Structure of TELODIN. FBW7 F-box and WD40 domains with the TELODIN motif colored in purple (PDB: 2OVP) (A), TELODIN in FBW7 WD40 domain (B), and synthetic TELODIN equilibrated conformation under a 300 ns MD simulation (C).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 Thermo Cat#A27034 Anti-rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 555 Thermo Cat#A27039 Anti-rabbit IgG-Peroxidase antibody produced in goat Sigma Cat#A6154 Anti-Rap1 antibody IMGENEX Cat#IMG-289 Anti-SP-C antibody (for western blot) Abcam Cat#ab90716 Anti-SP-C antibody (for immunofluorescence) Santa Cruz Biotechnology Cat#sc-7706 Anti-T1a antibody Biolegend Cat#127408 Anti-Ter119 antibody Biolegend Cat#116204 Anti-TIN2 antibody GeneTex Cat59B388 Anti-TPP1 antibody (for immunofluorescence) Novus Cat#NBP2-45475 Anti-TPP1 antibody (for western blot) Abcam Cat#ab112050 and ab54685 Anti-TRF1 antibody Cell Signaling Technology Cat#3529 Anti-TRF2 antibody Novus Cat#NB100-56506 Anti-a-SMA antibody Sigma Cat#A2547 Anti-a-Tubulin antibody Cell Signaling Technology Cat#2144 Peroxidase-AffiniPure goat anti mouse IgG Jackson Cat#115-035-062 Bacterial and Virus Strains Cre lentivirus Genomeditech Cat#GM-0220CR02 E. coli (DH5a) TransGen Biotech Cat#CD201-01 pEF-1a-TPP1 lentivirus GenePharma N/A pLMCS-FBW7a lentivirus Invitrogen (Shanghai) N/A pLMCS-FBW7g lentivirus Invitrogen (Shanghai) N/A Chemicals, Peptides, and Recombinant Proteins Bleomycin Sigma Cat#5507 BrdU Sigma Cat#B5002 CHIR99021 Sigma Cat#SML1046 Colcemid Thermo Cat# 15212012 Cycloheximide Sigma Cat#239763 Digoxin Roche Cat#11585614910 Dispase II Sigma Cat#4942078001 DMSO Sigma Cat#D8418 Dynabeads MyOne streptavidin T1 magnetic beads Thermo Cat#65601 Flag-coupled magnetic beads Sigma Cat#M8823 Glutathione-Sepharose 4B Sigma Cat#GE17-0756-01 H2O2 (3%) Sigma Cat#88597 Lipofectamine 2000 Thermo Cat#11668019 Masson Trichrome Sigma Cat#HT153 MG132 Sigma Cat#M8699 Peptides KE BIOCHEM See Table S5 PNA probe Panagene Cat#F1001,F1002 Streptavidin-PE Biolegend Cat#405204 TC-G24 TOCRIS Cat#4353 (Continued on next page) e2 Cell Metabolism 32, 1–18.e1–e9, November 3, 2020

Techniques: Binding Assay