Journal: Molecular Medicine Reports

Article Title: Pan-cancer analysis of the carcinogenic role of WSB2 in human tumors

doi: 10.3892/mmr.2025.13625

Figure Lengend Snippet: WSB2 promotes the proliferation and migration of breast cancer cells. (A) Reverse transcription-quantitative PCR and (B) western blotting were used to detect the effect of WSB2 overexpression in MDA-MB-231 and MCF-7 cells. (C) Viability of MDA-MB-231 and MCF-7 cells was evaluated using a CCK-8 assay. (D) Representative images and (E) the quantification of the number of MDA-MB-231 and MCF-7 cells in the S phase evaluated using an 5 ethynyl-2′-deoxyuridine assay. (F) Representative images of colony formation ability of MDA-MB-231 and MCF-7cells. (G) Representative images and (H) quantification of wound healing and (I) colony formation ability of MDA-MB-231 and MCF-7cells. (J) Transwell assays to determine the migration ability of MDA-MB-231and MCF-7cells. (K) ERK1/2, p53 and PCNA and (L) Snail, E-cadherin and Vimentin protein levels in MDA-MB-231 and MCF-7 cells were detected using western blotting. *P<0.05, **P<0.01, ***P<0.001. WSB2, WD repeat and SOCS box containing 2.

Article Snippet: Standard technique was used to carry out western blotting ( ) and the following specific primary antibodies were used: Anti-WSB2 (cat. no. ab127176; Abcam, 1:2,000), anti-GADPH (cat. no. CSB-MA000071Mom; Cusabio Technology, LLC, 1:10,000), anti-β-Actin (cat. no. AC026; ABclonal Biotech Co., Ltd. 1:10,000 dilution), anti-HA-tag (cat. no. B1021; Suzhou Botelon Immunotechnology Co., Ltd. 1:5,000), anti-E-cadherin (cat. no. BD-PT1454; Suzhou Botelon Immunotechnology Co., Ltd.; 1:500 dilution), anti-proliferating cell nuclear antigen (PCNA; cat. no. D220014-0025; Sangon Biotech Co., Ltd. 1:1,000 dilution), anti-Vimentin (cat. no. BD-PB4686; Suzhou Botelon Immunotechnology Co., Ltd. 1:500), anti-p53 antibody (p53) (cat. no. CSB-PA07889A0Rb; Cusabio Technology, LLC.

Techniques: Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, CCK-8 Assay

Journal: BMC Biology

Article Title: m6A-modified circZNF548 regulates exosomal miR-7108-3p to activate CD3 + CD8 + T cells and suppress NSCLC growth by JMY

doi: 10.1186/s12915-025-02355-z

Figure Lengend Snippet: JMY is a target of miR-7108-3p. A The binding sites of JMY-3’-UTR (Mu-JMY with mutant site) and miR-7108-3p. B miR-7108-3p regulated luciferase expression in the luci-JMY-3’-UTR- or luci-Mu-JMY-3’-UTR-treated A549 or H1975 cells. Data are presented as the mean ± SD. **p < 0.01; Student’s t-test. ns, no significance. C Immunoblotting analyzing the expression of JMY and p53-related apoptotic genes in A549 cells. D - F JMY levels in miR-7108-3p-treated and circZNF548-treated cells was detected by immunofluorescence. Bar = 125 μm. G JMY-3’-UTR recovery affected the role of miR-7108-3p and inhibitor miR-7108-3p in cell growth. Data are presented as the mean ± SD. **p < 0.01; ANOVA. H , I JMY recovery affected the role of miR-7108-3p and inhibitor miR-7108-3p in cell migration, respectively. bar = 100 μm. Data are presented as the mean ± SD. **p < 0.01; ANOVA. J Immunohistochemical staining showed JMY levels increased in adjacent tissues. The score of staining was shown below the figure. bar = 125 μm. Data were present as median (interquartile range), **p < 0.01; Mann–Whitney U test. K qRT-PCR analysis showed that JMY expression was low in NSCLC tissues. Data were present as median (interquartile range), **p < 0.01; Mann–Whitney U test. L The relationship between JMY expression and miR-7108-3p levels. M The relationship between JMY and circZNF548 levels. N Overall survival analysis of JMY expression in patients with NSCLC

Article Snippet: Membranes were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-human GAPDH (1:3000, BS65656, Bioworld, MN, USA), rabbit anti-human P53 (1:1000, 60,283–2-lg, Proteintech, PA, USA), rabbit anti-human JMY (1:500, 25,098–1-AP, Proteintech), rabbit anti-human Bax (1:500, BS79706, Bioworld), rabbit anti-human Bcl2 (1:500, BZ16777, Bioworld), rabbit anti-human CD8a (1:500, 48,750–1, Signalway Antibody, MD, USA).

Techniques: Binding Assay, Mutagenesis, Luciferase, Expressing, Western Blot, Immunofluorescence, Migration, Immunohistochemical staining, Staining, MANN-WHITNEY, Quantitative RT-PCR

Journal: BMC Biology

Article Title: m6A-modified circZNF548 regulates exosomal miR-7108-3p to activate CD3 + CD8 + T cells and suppress NSCLC growth by JMY

doi: 10.1186/s12915-025-02355-z

Figure Lengend Snippet: Exosomal miR-7108-3p regulated cancer cell growth in NSG mice. A - C Tumor size, volume changes, and weights of xenografts were analyzed in exosomal miR-7108-3p-treated group compared with controls. N = 5. D , E The PRF1 and Gzmb levels in the sera of exosomal miR-7108-3p-treated mice. F CD8 immunofluorescence analysis in xenografts. Bar = 125 μm. G Proposed model by which circZNF548 suppressed NSCLC proliferation by regulating JMY-p53 related genes and promoted the killing function of CTL through CD8. Data were shown as median (interquartile range), **p < 0.01; Student’s t-test or Mann–Whitney U test

Article Snippet: Membranes were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-human GAPDH (1:3000, BS65656, Bioworld, MN, USA), rabbit anti-human P53 (1:1000, 60,283–2-lg, Proteintech, PA, USA), rabbit anti-human JMY (1:500, 25,098–1-AP, Proteintech), rabbit anti-human Bax (1:500, BS79706, Bioworld), rabbit anti-human Bcl2 (1:500, BZ16777, Bioworld), rabbit anti-human CD8a (1:500, 48,750–1, Signalway Antibody, MD, USA).

Techniques: Immunofluorescence, MANN-WHITNEY

Journal: NPJ Vaccines

Article Title: Oncolytic virus-mediated p53 activation boosts the antitumor immunity of a p53-transduced dendritic cell vaccine

doi: 10.1038/s41541-025-01219-5

Figure Lengend Snippet: A Schematic illustration of the experimental protocol of preparation of dendritic cells (DCs) transduced with the replication-deficient, wild-type p53-expressing adenovirus Ad-p53 (Ad-p53 DCs). Bone marrow-derived cells isolated from the femur of BALB/c mice were incubated with GM-CSF (40 ng/mL) and IL-4 (10 ng/mL) for 5 days and then infected with Ad-p53 (100 MOI) for 2 days to obtain Ad-p53 DCs. Figures were generated using BioRender. B Expression of human p53 mRNA in control DCs and DCs infected with DL312 (100 MOI) or Ad-p53 (100 MOI) for 48 h. GAPDH was used as a control gene. C Representative photographs of expression of human p53 protein in DCs infected with Ad-p53 (100 MOI) for 72 h. Scale bars, 100 μm. D Expression of mature DC markers in Ad-p53 DCs. Control DCs and Ad-p53 DCs were subjected to flow cytometry for analysis of CD11c, CD86, MHC class-II (MHC-II), CD103, and CCR7 expression. The mean fluorescence intensity (MFI) for each protein was determined by calculating the difference between the MFI of the antibody- and control IgG-treated cells. Data are expressed as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s comparison test. *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were placed on slides and stained overnight with the primary antibody against human p53 (1:800, 2527; Cell Signaling Technology, Beverly, MA, USA).

Techniques: Transduction, Expressing, Derivative Assay, Isolation, Incubation, Infection, Generated, Control, Flow Cytometry, Fluorescence, Comparison

Journal: NPJ Vaccines

Article Title: Oncolytic virus-mediated p53 activation boosts the antitumor immunity of a p53-transduced dendritic cell vaccine

doi: 10.1038/s41541-025-01219-5

Figure Lengend Snippet: A Viability of CT26 cells was assessed using an XTT assay 3 days after OBP-702 treatment at the indicated MOIs. Cell viability was calculated relative to that of mock-infected cells, which were set as 1.0. B Expression of human p53 mRNA in CT26 cells before and after infection with OBP-702 (100 MOI) for 24 h. Data are expressed as mean ± SD ( n = 3). Statistical significance was determined using the Student t -test. **** P < 0.0001. C Whole-cell lysates of CT26 cells before and after infection with OBP-702 (100 MOI) for 24 h were subjected to Western blot analysis of mouse p53, human p53, and β-actin expression. D Schematic illustration of the CT26 tumor model experimental protocol. Figures were generated using BioRender. E Photographs of tumors in each group. F Growth curves for control, Ad-p53 DC-treated, OBP-702-treated, and combination therapy (combo)-treated tumors. G Weight of control, Ad-p53 DC-treated, OBP-702-treated, and combo-treated tumors. Data are expressed as the mean ± SD ( n = 5). Statistical significance was determined using one-way ANOVA followed by Tukey’s comparison test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Cells were placed on slides and stained overnight with the primary antibody against human p53 (1:800, 2527; Cell Signaling Technology, Beverly, MA, USA).

Techniques: XTT Assay, Infection, Expressing, Western Blot, Generated, Control, Comparison

Journal: NPJ Vaccines

Article Title: Oncolytic virus-mediated p53 activation boosts the antitumor immunity of a p53-transduced dendritic cell vaccine

doi: 10.1038/s41541-025-01219-5

Figure Lengend Snippet: A Combined effect of Ad-p53 DC and OBP-702 in the CT26 tumor model using athymic nude mice. Data are expressed as the mean tumor volume ± SD ( n = 5). B Combined effect of Ad-p53 DC and OBP-301 in the CT26 tumor model. Data are expressed as the mean tumor volume ± SD ( n = 6). C Difference in the growth of CT26 tumors between control and complete response (CR) mice. Combination therapy-treated CR mice and control mice were subcutaneously inoculated with CT26 cells. Data are expressed as the mean ± SD ( n = 3). D Growth curves for treated and untreated sites of control, Ad-p53 DC-treated, OBP-702-treated, and combination therapy (combo)-treated tumors in bilateral CT26 tumor models. Data are expressed as the mean ± SD ( n = 5). E Representative photographs of immunohistochemical staining for CD8 + T cells and CD11c+ DCs in each group. Scale bars, 50 μm. F Percentage of CD8+ cells and CD11c+ cells calculated from five different randomly selected fields. Data are expressed as the mean ± SD ( n = 5). Statistical significance was determined using one-way ANOVA followed by Tukey’s comparison test. * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: Cells were placed on slides and stained overnight with the primary antibody against human p53 (1:800, 2527; Cell Signaling Technology, Beverly, MA, USA).

Techniques: Control, Immunohistochemical staining, Staining, Comparison

Journal: NPJ Vaccines

Article Title: Oncolytic virus-mediated p53 activation boosts the antitumor immunity of a p53-transduced dendritic cell vaccine

doi: 10.1038/s41541-025-01219-5

Figure Lengend Snippet: A Splenocytes isolated from control and Ad-p53 DC-immunized mice were stimulated with IL-2 and co-cultured with OBP-702-infected and non-infected CT26 cells for 6 h and then subjected to FACS analysis. The percentage of IFN-γ+ and granzyme B+ cells among CD8+ cells was assessed by flow cytometry. Data are expressed as the mean ± SD ( n = 3 or 4). B MHC-II-bound peptides were isolated from control, DL312-infected, and Ad-p53-infected DCs. MHC-I-bound peptides were isolated from control, OBP-301-infected, and OBP-702-infected CT26 cells. Human p53-derived p53 63-79 peptide (APRMPEAAPPVAPAPAA) was commonly expressed in the context of MHC-I and MHC-II molecules. Figures were generated using BioRender. C Splenocytes isolated from control and Ad-p53 DC-immunized mice were incubated with p53 peptide for 72 h and subjected to ELISPOT assay. The number of spots per 1 × 10 6 splenocytes is shown. Figures were generated using BioRender. Data are expressed as the mean ± SD ( n = 3). D Splenocytes isolated from control and Ad-p53 DC-immunized mice were incubated with p53 peptide and incubated for 72 h. The percentage of IFN-γ+ and granzyme B+ cells among CD8+ cells was assessed by flow cytometry. Data are expressed as the mean ± SD ( n = 3). The statistical significance of differences between two groups or four groups was determined using the Student t -test or one-way ANOVA followed by Tukey’s comparison test, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Cells were placed on slides and stained overnight with the primary antibody against human p53 (1:800, 2527; Cell Signaling Technology, Beverly, MA, USA).

Techniques: Isolation, Control, Cell Culture, Infection, Flow Cytometry, Derivative Assay, Generated, Incubation, Enzyme-linked Immunospot, Comparison

Journal: NPJ Vaccines

Article Title: Oncolytic virus-mediated p53 activation boosts the antitumor immunity of a p53-transduced dendritic cell vaccine

doi: 10.1038/s41541-025-01219-5

Figure Lengend Snippet: A Viability of MC38 cells was assessed using an XTT assay 3 days after OBP-702 treatment at the indicated MOIs. Cell viability was calculated relative to that of mock-infected cells, which were set as 1.0. B Expression of human p53 mRNA in MC38 cells before and after infection with OBP-702 (100 MOI) for 24 h. Data are expressed as mean ± SD ( n = 3). C Whole-cell lysates of MC38 cells before and after infection with OBP-702 (100 MOI) for 24 h were subjected to Western blot analysis of mouse p53, human p53, and β-actin expression. D Growth curves of control and Ad-p53 DC-treated tumors. Data are expressed as mean ± SD ( n = 3). E Schematic illustration of the MC38 tumor model experimental protocol. Figures were generated using BioRender. F Photographs of tumors in each group. G Growth curves of control, Ad-p53 DC-treated, OBP-702-treated, and combination therapy (combo)-treated tumors. H Representative photographs of immunohistochemical staining for CD8 + T cells and CD11c+ DCs in each group. Scale bars, 50 μm. I Percentage of CD8+ cells and CD11c+ cells calculated from five different randomly selected fields. Data are expressed as the mean ± SD ( n = 5). J CD8 + T cells isolated from splenocytes of control and treated mice were co-cultured with MC38 cells for 24 h, and the amount of extracellular LDH released from tumor cells was determined. The statistical significance of differences between two groups or three groups was determined using the Student t -test or one-way ANOVA followed by Tukey’s comparison test, respectively. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: Cells were placed on slides and stained overnight with the primary antibody against human p53 (1:800, 2527; Cell Signaling Technology, Beverly, MA, USA).

Techniques: XTT Assay, Infection, Expressing, Western Blot, Control, Generated, Immunohistochemical staining, Staining, Isolation, Cell Culture, Comparison