Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 facilitates ASCC2 nuclear translocation and promotes ASCC2–ALKBH3 interaction. A . Silver staining (left panel) analysis of proteins isolated from AGS cells using ChIRP with DLEU1 or the control LacZ probes. The red rectangle highlighted the potential presence of ASCC2 in the DLEU1 complex, subsequently confirmed by LC/MS mass spectrometry analysis (right panel). B - C . RNA pull-down ( B ) and RIP assays ( C ) demonstrated the interaction between ASCC2 and DLEU1 in AGS and HGC27 cells. D . Immunofluorescence staining showed co-localization of DLEU1 with ASCC2, Scale bar = 10 μm. E . Western blot results showed that overexpression of DLEU1 increased ASCC2 protein levels in MGC803 and AGS cells, whereas knockdown of DLEU1 decreased ASCC2 protein levels in HGC27 and MKN45 cells. F - G . Nuclear-cytoplasmic fractions ( F ) and Immunofluorescence staining ( G ) demonstrate increased nuclear localization of ASCC2 in DLEU1 overexpressed cells. Scale bar = 10 μm. H . Endogenous co-immunoprecipitation assays reveal enhanced ASCC2–ALKBH3 binding following DLEU1 overexpression. I . Western blot analysis showed changes in ASCC2 and ALKBH3 protein expression in AGS with DLEU1 overexpression or combined DLEU1 overexpression with ASCC2 knockdown(left panel), and in HGC27 cells with DLEU1 knockdown or combined DLEU1 knockdown with ASCC2 overexpression (right panel). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. See also Figures

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Translocation Assay, Silver Staining, Isolation, Control, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Immunofluorescence, Staining, Western Blot, Over Expression, Knockdown, Immunoprecipitation, Binding Assay, Expressing

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 promotes G6PD transcription via ASCC2/ALKBH3-mediated recruitment of E2F1. A - B . qRT-PCR and Western blot analysis of E2F1 expression in AGS and MGC803 cells upon ALKBH3 overexpression. C . Time-course analysis of E2F1 mRNA levels in AGS and MGC803 cells transfected with vector or ALKBH3. D - E . RT-qPCR and western blot analyzed of E2F1 and G6PD expression in AGS and MGC803 cells following DLEU1 overexpression or DLEU1 overexpression combined with ASCC2 knockdown. F - G . RT-qPCR and western blot analyzed of G6PD levels in AGS and MGC803 cells with E2F1 overexpression. H . qRT-PCR analysis of G6PD mRNA expression in AGS and MGC803 cells after ALKBH3 overexpression or co-transfection with ALKBH3 and shRNA targeting E2F1 (shE2F1). I . Western blot analysis of ASCC2, G6PD and E2F1 expression in AGS and MGC803 cells with ALKBH3 overexpression or ALKBH3 overexpression combined with E2F1 knockdown. J . ChIP-qPCR analysis confirmed E2F1 binding to the G6PD promoter and intron 2 regions in AGS cells. K . Luciferase reporter assay showed the transcriptional activity of wild-type and mutant G6PD promoter and intron 2 regions in AGS cells. L . Luciferase assays demonstrate that knockdown of DLEU1 , ASCC2, or ALKBH3 attenuates E2F1-mediated activation of G6PD transcription. L - M . ChIP-qPCR and luciferase reporter validated E2F1 binding to an intronic region of ASCC2 in AGS cells. M . Luciferase assays indicate that knockdown of DLEU1 , ASCC2, or ALKBH3 reduces E2F1-driven ASCC2 transcription. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Transfection, Plasmid Preparation, Knockdown, Cotransfection, shRNA, ChIP-qPCR, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Mutagenesis, Activation Assay

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 knockdown suppresses tumor growth by targeting ASCC2 and G6PD in xenograft models. A . Schematic representation of the xenograft experiment. MKN45 cells with stable knockdown of DLEU1 ( shDLEU1 ) or co-knockdown of DLEU1 and ASCC2 ( shDLEU1 + shASCC2 ) were subcutaneously implanted into nude mice. G6PD inhibitor (G6PDi-1) was administered intraperitoneally (10 mg/kg, every 2 days, for a total of six doses) from day 15 to day 21. B . Tumor growth analysis across different groups. Left: Representative images of tumors from the indicated groups. Middle: Tumor weight comparison among groups. Right: Tumor volume progression over time. C . IHC staining for ASCC2, ALKBH3, E2F1, and G6PD in tumor tissues. Representative IHC images (left) and quantification of staining scores (right) for each marker. Scale bar = 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Knockdown, Comparison, Immunohistochemistry, Staining, Marker

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: Expression and correlation analysis of DLEU1 and associated markers in normal and GC tissues, and its epigenetic regulation in GC. A . Representative multiplex immunofluorescence staining images showing DLEU1 (green), ASCC2 (white), ALKBH3 (orange), and E2F1 (cyan) in normal and tumor tissues ( N = 26, T = 104). Scale bar = 50 μm. B . Quantification of DLEU1 -positive cell density in normal and tumor tissues. C . Quantification of ASCC2-, ALKBH3-, and E2F1-positive cell densities in normal and tumor tissues. D . Percentage of E2F1 localization in the nucleus and cytoplasm in normal and tumor tissues. E . Scatter plots showing positive correlations between DLEU1 expression and ASCC2, ALKBH3, and E2F1 expression in tumor tissues. F . Correlation between DLEU1 expression and E2F1 localization in the cytoplasm and nucleus in tumor tissues. G . Correlation between ASCC2- and ALKBH3-positive cell in tumor tissues. H . HGC27 and MKN45 cells were treated with DMSO (control), Etoposide (10 µM, 24 h), 5-Fluorouracil (5-FU, 20 µg/mL, 24 h), or H 2 O 2 (10 µM, 2 h). The enrichment of H3K4me3 and H3K27ac at the DLEU1 promoter was analyzed by ChIP-qPCR. I . Proposed model illustrating how DLEU1 promotes gastric cancer progression via the ASCC2/ALKBH3/E2F1/G6PD axis, impacting transcriptional activation, DNA repair, and redox homeostasis. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Expressing, Multiplex Assay, Immunofluorescence, Staining, Control, ChIP-qPCR, Activation Assay

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: DEPP1 is aberrantly overexpressed in gastric cancer. (A) Venn diagrams illustrating the differentially expressed genes. (B) Expression levels of DEPP1 in GC and normal tissues in the TCGA-STAD cohort. Relative expression of DEPP1 in 11 pairs of GC and adjacent normal tissues measured by (C) reverse transcription-quantitative PCR and (D) western blotting. (E) Representative immunofluorescence images of DEPP1 in 11 pairs of GC and adjacent normal tissues. *P<0.05, **P<0.01 and ***P<0.001. DEPP1, decidual protein induced by progesterone; GC, gastric cancer; TCGA-STAD, The Cancer Genome Atlas Stomach Adenocarcinoma; N, normal; T, tumor; Ns, not significant.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: Prognostic value of DEPP1 is explored in the TCGA-STAD cohort. Kaplan-Meier survival analyses of DEPP1 in patients with GC on (A) overall survival and (B) disease-free survival. DEPP1 expression between patients with GC who are (C) alive or dead and (D) patients with or without disease progression or recurrence. (E) Univariate and (F) multivariate Cox regression analysis of DEPP1 and other clinicopathologic features. (G) Receiver operating characteristic curves of DEPP1 and conventional oncogenes EGFR and KRAS in predicting five-year overall survival of patients with GC. **P<0.01 and ***P<0.001. DEPP1, decidual protein induced by progesterone; TCGA-STAD, The Cancer Genome Atlas Stomach Adenocarcinoma; GC, gastric cancer; OS, overall survival; DFS, disease-free survival; HR, hazard ratio; CI, confidence interval.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing, Biomarker Discovery

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: Prognostic value of DEPP1 is validated in GSE84437 , GSE15459 and Renji cohorts. Kaplan-Meier survival analyses of DEPP1 on overall survival in (A) GSE84437 and (D) GSE15459 cohorts. (B) Univariate and (C) multivariate Cox regression analysis of DEPP1 and other clinicopathologic parameters in the GSE84437 cohort. (E) Univariate and (F) multivariate Cox regression analysis of DEPP1 and other clinicopathologic factors in the GSE15459 cohort. (G) Representative immunohistochemistry images of DEPP1 in gastric cancer and normal tissues. (H) Kaplan-Meier survival analysis of DEPP1 on overall survival in the Renji cohort. DEPP1, decidual protein induced by progesterone; OS, overall survival; DFS, disease-free survival; HR, hazard ratio; CI, confidence interval.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Immunohistochemistry

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: Establishment and evaluation of a prognostic nomogram for GC. (A) Construction of a nomogram integrating DEPP1, age and TNM stage to predict the 1-, 3- and 5-year overall survival of patients with GC. (B) Calibration curves to assess the predictive accuracy of the nomogram for survival outcomes. (C) Results of decision curve analysis comparing the nomogram's performance with that of individual predictors, including DEPP1, age and TNM stage. DEPP1, decidual protein induced by progesterone; GC, gastric cancer; TNM, tumor-node-metastasis; OS, overall survival; Pr, probability.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques:

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: Relationship between DEPP1 expression and clinicopathologic factors in patients with GC. (A) DEPP1 expression levels across different Lauren phenotypes. (B) Kaplan-Meier survival analyses of DEPP1 in diffuse, intestinal and mixed subtypes of the GSE15459 cohort. (C) DEPP1 expression levels in stage I and advanced stages of GC. (D) Kaplan-Meier survival analyses of DEPP1 in stage I, II, III and IV of the TCGA-STAD cohort. (E) DEPP1 expression levels across distinct T stages. (F) The histogram displaying the distribution of T stages between the low- and high-expression groups of DEPP1. (G) The alluvial diagram depicting the association between DEPP1 expression, T stages and overall survival status. (H) Kaplan-Meier survival analyses of DEPP1 in T1, T2, T3 and T4 stages of the GSE84437 cohort. DEPP1, decidual protein induced by progesterone; GC, gastric cancer; TCGA-STAD, The Cancer Genome Atlas Stomach Adenocarcinoma.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: DEPP1 correlates with a stroma-rich and immunosuppressive microenvironment of GC. (A) The stromal, immune, and estimate scores in the low- and high-expression groups of DEPP1. (B) The correlation between DEPP1 expression and stromal score. (C) The distribution of various immune and stromal cells in the GC microenvironment estimated by the EPIC algorithm. (D) The heatmap showing the correlation between DEPP1 expression and immune and stromal cell populations. The infiltration levels of distinct immune and stromal cells in the low- and high-expression groups of DEPP1, as assessed by (E) EPIC, (F) TIMER and (G) quanTIseq methods, respectively. (H) A correlation heatmap illustrating the relationships between DEPP1 expression and various immune and stromal cells based on the results of the quanTIseq algorithm. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. DEPP1, decidual protein induced by progesterone; GC, gastric cancer; NK, natural killer; ns, not significant.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: Potential regulatory mechanisms underlying DEPP1. (A) The volcano plot depicting the differentially expressed genes between the low- and high-expression groups of DEPP1. (B) Gene Ontology and (C) Kyoto Encyclopedia of Genes and Genome enrichment analyses of the differentially expressed genes. Gene Set Enrichment Analysis demonstrates the hallmark pathways enriched in the (D) high- and (E) low-expression groups of DEPP1. (F) The box plot of the downstream hub genes of DEPP1. (G) The correlation heatmap displays the relationships among the hub genes. DEPP1, decidual protein induced by progesterone.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: High DEPP1 expression predicts poor chemotherapy sensitivity in patients with GC. (A) The association between DEPP1 expression levels and the IC 50 of selective chemotherapy drugs as shown by the heatmap. The estimated IC 50 of chemotherapy drugs in the low- and high-expression groups of DEPP1, as analyzed using data from the (B) GDSC and (C) CTRP datasets, respectively. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. DEPP1, decidual protein induced by progesterone; GC, gastric cancer; IC 50 , half-maximal inhibitory concentration; GDSC, Genomics of Drug Sensitivity in Cancer; CTRP, Cancer Therapeutics Response Portal; ns, not significant.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: Expressing, Concentration Assay

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: DEPP1 enhances oxaliplatin resistance in gastric cancer cells in vitro . Effects of oxaliplatin and fluorouracil supplementation on DEPP1 protein levels in (A) MKN45 and (B) HGC27 cells, respectively. Validation of DEPP1 overexpression in (C) MKN45 and (D) HGC27 cells. (E) Impact of DEPP1 overexpression on MKN45 cell proliferation, assessed using EdU flow cytometry. (F) Influence of DEPP1 overexpression on MKN45 proliferation following a 24-h treatment with 10 µM oxaliplatin. (G) Effects of ectopic DEPP1 expression on oxaliplatin-induced apoptosis (20 µM) in MKN45 cells, as measured by Annexin V/PI flow cytometry. (H) Analysis of full-length and cleaved PARP protein levels following forced DEPP1 expression in (H) MKN45 and (J) HGC27 cells. *P<0.05 and ***P<0.001 DEPP1, decidual protein induced by progesterone. ns, not significant.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: In Vitro, Biomarker Discovery, Over Expression, Flow Cytometry, Expressing

Journal: Oncology Reports

Article Title: DEPP1: A prognostic biomarker linked to stroma-rich and immunosuppressive microenvironment, promoting oxaliplatin resistance in gastric cancer

doi: 10.3892/or.2025.8915

Figure Lengend Snippet: DEPP1 enhances oxaliplatin resistance in gastric cancer cells in vivo . (A) Schematic representation of the subcutaneous mouse model receiving either 5% GS or 5 mg/kg oxaliplatin intraperitoneally. (B) Images and (C) weights of control and DEPP1-overexpressing tumors treated with or without oxaliplatin. (D) Representative immunohistochemistry images of DEPP1, Ki67, and cleaved caspase 3 in subcutaneous tumor tissues. DEPP1, decidual protein induced by progesterone; GS, glucose saline.

Article Snippet: The membranes were then incubated with 5% skimmed milk at room temperature for 1 h. Following this, the membranes were probed with primary antibodies against DEPP1 (1:1,000; cat. no. 25833-1-AP; Proteintech Group, Inc.), PARP (1:1,000; cat. no. 9532; Cell Signaling Technology, Inc.), cleaved PARP (1:500; cat. no. HY- P80448 ; MedChemExpress), GAPDH (1:1,000; cat. no. 5174; Cell Signaling Technology, Inc.), or β-actin (1:20,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) at 4°C overnight, followed by incubation with peroxidase-conjugated goat anti-rabbit IgG (33101ES60, 1:5,000, Shanghai Yeasen Biotechnology Co., Ltd.) or peroxidase-conjugated goat anti-mouse IgG (1:5,000; cat. no. BL001A; Biosharp Life Sciences) at room temperature for 1 h. Finally, proteins were visualized using an Omni-ECLTMPico Light Chemiluminescence Kit (cat. no. SQ202L; Epizyme, Inc.).

Techniques: In Vivo, Control, Immunohistochemistry, Saline