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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.

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

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 (****).

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

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 (****).

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