Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 1 High expression of TRAF7 is closely associated with poor prognosis in glioma. A The expression of TRAF7 mRNA was quantitatively analyzed between the glioma tumor site and normal brain tissues based on TCGA database. B-C Kaplan–Meier survival and ROC curves using TCGA. D-E Kaplan–Meier survival curves and ROC for predicting the 1-, 3- and 5-year overall survival (p < 0.0001) in CCGA. F The Nomogram model was built to analyze prognostic factors in 1-, 3-, and 5-year OS of glioma patients. G Decision curve analysis (DCA) for the clinical benefits and application of the nomogram. H-J Calibration curves of the nomogram model for 1-year, 3-year, and 5-year OS. K TRAF7 expression were examined by qRT-PCR in six glioma cell lines. L The knockdown efficiency of TRAF7 mRNA in Hs683 and U251 cells. M The protein expression of TRAF7 was conducted by western blot. N–O TRAF7 protein expression was quantified in Hs683 and U251 cells. Data are presented as the mean ± SD

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 2 The clinical characteristics of glioma patients. A Heat map of 143 glioma patients with clinical characteristics, including 76 recurrent samples. B-D The TRAF7 expression levels of glioma patients displayed differences in their pathological stages. E–F The representative images of Hematoxylin and IHC staining were shown in different TRAF7 expression groups (The scale bar for HE staining and 40 × IHC staining is 50 µm, and the scale bar for 20 × IHC staining is 100 µm). G-H Survival curves of RFS and OS glioma patients with varying levels of TRAF7 expression

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 3 The knockdown of TRAF7 inhibits cell proliferation and migration in vitro. A-B Transwell assays in Hs683 and U251 cells. C-D Colony formation assays visualized on day 14. E–F The Hs683 and U251 cell lines with TRAF7 knockdown significantly inhibited cell proliferation by EDU assays. G-I Wound healing assays and migration rate at 24 and 48 h. J-K Cell viability was determined by the CCK-8 assay. Data are presented as the mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Knockdown, Migration, In Vitro, CCK-8 Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 5 Combination of Lomustine (CCNU) and sh-TRAF7 promotes glioma senescence and G0/G1 arrest. A IC50 of lomustine (CCNU). B Cell viability of TRAF7 deficient cell lines treated with low concentrations of lomustine (1 μM and 10 μM) for 12 h. C Cell viability of Hs683 and U251 cells treated with lomustine (50 μM). D SA-β-gal staining after CCNU treatment of the control group and sh-TRAF7 group in the cells. E–F The mRNA expression and protein expression of G0/G1 arrest related genes after CCNU treatment. G-H mRNA expression and protein expression of senescence related genes after CCNU treatment in the cells

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Staining, Control, Expressing

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 6 TRAF7 depletion inhibits glioma proliferation and induces senescence via KLF4. A Kaplan–Meier survival using TCGA on KLF4. B The overexpression of KLF4 in Hs683 and U251 cells. C The expression of TRAF7 and KLF4 in the Hs683 and U251 glioma cell lines. D Co-immunoprecipitation (Co-IP) assays were performed using lysates from U251 cells transfected with Flag-tagged KLF4 and HA-tagged TRAF7. E–F Colony formation assays were performed in the glioma cell lines with or without KLF4 overexpression. G-H SA-β-gal staining was performed in the glioma cell lines (scale bar = 50 µm). I-J The rescue colony formation assay showed that the inhibitory effects of TRAF7 knockdown on cell proliferation could be rescued by additional KLF4 overexpression in cells. K-L The rescue SA-β-gal staining assay showed that the promotion effects of TRAF7 knockdown on cell senescence could be rescued by additional KLF4 overexpression in cells (scale bar = 50 µm). *p < 0.05, **p < 0.01, and ***p < 0.001

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Over Expression, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Staining, Colony Assay, Knockdown

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 7 Establishment of patient-derived primary and recurrent glioma stem cell spheres (GSCs). A The expression of TRAF7 in primary and recurrent patient-derived glioma cells with or without knockdown. B IC50 of Lomustine (CCNU) in primary and recurrent glioma stem cells. C Cell viability of primary and recurrent patient-derived glioma cells treated with lomustine (50 μM). D Brightfield image of GSC spheres in Matrigel (scale bar = 100 µm). E The radar chart provides a comprehensive comparison of different cells, including the inhibitory efficiency of combined therapy, tumor sphere size, CD133 expression levels, and migration capacity. F Evaluation of the GSCs formation ability between the control group and si-TRAF7 group treated with or without CCNU (scale bar = 100 µm). G The primary and recurrent glioma stem cells were stained with stem cell markers (CD133) (scale bar = 100 µm). H Transwell assays of pGSC#3 and rGSC#1 (scale bar = 50 µm). I Heat map of three primary glioma patients and one recurrent glioma patient with clinical characteristics

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Derivative Assay, Expressing, Knockdown, Comparison, Migration, Control, Staining

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 8 Sh-TRAF7 and Lomustine (CCNU) synergistically inhibit glioma tumor growth. A Schematic illustration of the experimental design for the glioma orthotopic implantation model, and evaluation of brain tumor growth after injection CCNU. B Bioluminescence images of nude mice bearing glioma orthotopic xenograft (n = 5) were shown on days 14, 21, 28, 35, and 42. The nude mice were inoculated with U251 cells transfected with luciferase, divided into six groups: (1) Control, (2) Control + CCNU, (3) sh-TRAF7, (4) sh-TRAF7 + CCNU, (5) OE-TRAF7, (6) OE-TRAF7 + CCNU. The crossing in the blank area indicated that the corresponding mouse had died. C-D Survival curves of glioma-bearing mice, with median survival times listed in the accompanying table. E Quantitative analysis of bioluminescence. F The curves of body weight in nude mice

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Injection, Transfection, Luciferase, Control

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 9 Histological analyses of TRAF7 inhibition and lomustine via mediating cellular senescence and G0/G1 arrest in vivo. A The whole brain of glioma nude mice were stained HE and TRAF7 (Full scan: scale bar = 2.5 mm; higher-magnification part below: scale bar = 100 µm). B Multiplex immunofluorescence staining image of cellular senescence markers (P21 and P53) and G0/G1 arrest markers (CCND1) and Ki67 in the tumor tissue. C IHC staining the cellular senescence markers (P21 and P53) and Ki67 in tumor tissue sections (scale bar = 100 µm)

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: Inhibition, In Vivo, Staining, Multiplex Assay, Immunofluorescence, Immunohistochemistry

Journal: Journal of experimental & clinical cancer research : CR

Article Title: TRAF7 knockdown induces cellular senescence and synergizes with lomustine to inhibit glioma progression and recurrence.

doi: 10.1186/s13046-025-03363-1

Figure Lengend Snippet: Fig. 10 A schematic mechanism overview of sh-TRAF7 and lomustine via mediating cellular senescence and G0/G1 arrest synergistic inhibit glioma tumor growth

Article Snippet: The sections were maintained with primary antibodies at 4°C throughout the night: TRAF7 (1:300, 11,780–1-AP, Proteintech, China), Ki67 (1:400, #9449, CST, USA), P53 (1:200, #2527, CST, USA), P21 (1:100, #2947, CST, USA), CCND1 (1:200, ab134175, Abcam, UK), CDK2 (1:100, ab32147, Abcam, UK).

Techniques: