Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: Specific IFN-γ and TNF-α release of T lymphocytes transduced with TIM-3-silenced HER2-specific chimeric antigen receptor (CAR) or HER2-specific CAR. (A, B) TIM-3-silenced CAR-T cells and control T cells were co-incubated with Galectin-9 + or Galectin-9 – SKOV3 tumor cells (E:T ratio 5:1 or 10:1). At 20 h after coculture, a specific enzyme-linked immunosorbent assay was used to analyze the supernatant for IFN-γ cytokine-release. Results were presented as mean ± standard deviation. (C, D) The detection of TNF-α in the same culture supernatant. Results were presented as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Transduction, Control, Incubation, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: TIM-3 silencing augmented the anti-tumor activity of chimeric antigen receptor-T (CAR-T) cells in vivo . 2 × 10 6 SKOV3 tumor cells expressing luciferase were intraperitoneally inoculated in a xenograft mouse model, and 7 days after inoculation, the 2 × 10 6 HER2-specific CAR-T kdTim-3 cells or CAR-T cells, or untreated T cells were intraperitoneally administered. (A, B) Tumor growth was monitored using an in vivo imaging system. (C) Survival curve of 80-day post-treatment. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Activity Assay, In Vivo, Expressing, Luciferase, In Vivo Imaging

Journal: Journal of Cell Science

Article Title: Interplay between nuclear survivin and the PRC2 complex and its impact on H3K27me3-directed transcriptional repression

doi: 10.1242/jcs.264572

Figure Lengend Snippet: Hypoxia increases expression of EZH2, H3K27me3 and survivin. (A) Immunoblots of WCEs from U2OS, HeLa and MRC5 lines cultured under normoxic or hypoxic environments (24 h). Blots were immunoprobed with anti-EZH2, anti-H3K27me3 and anti-survivin antibodies. Anti-Hif1a used to prove the hypoxic state had been induced, and anti-tubulin was used as a loading control. (B–D) Quantification of immunoblots represented in A from three independent experiments demonstrating that EZH2, H3K27me3 and survivin are all more abundant under hypoxia. Data presented are means±s.d. * P <0.05, ** P <0.01, *** P <0.001 (two-way ANOVA with Tukey's multiple comparisons post test).

Article Snippet: Human cervical cancer cells (HeLa), human bone osteosarcoma cells (U2OS), retinal pigment epithelial cells (RPE) and human breast cancer cells (MCF7) were originally from ATCC.

Techniques: Expressing, Western Blot, Cell Culture, Control

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH increases docetaxel efficacy against TNBC spheroids. MDA-MB-231 cells were plated in ultra-low attachment surface 24 well plates in complete medium and treated with increasing concentrations of docetaxel in the presence of DMSO (control, 0 PAH), PAH 10 µM or 30 µM. After 2 weeks pictures were taken using Cytation 5 cell imaging system from Biotek. The surface of spheroids was calculated and reported after normalization on the condition without docetaxel for each concentration of PAH.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Control, Imaging, Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH addition to chemotherapy inhibits migration of TNBC cells. A. 100000 cells were seeded on membrane from Transwell plates. Cells were treated 24 h with doxorubicin ± 15µM PAH or 48 h with docetaxel ± 15µM PAH. After fixation and staining with crystal violet, wells were observed on a microscope with X5 objective. B. Migration was measured using a wound-healing assay. A wound was performed on MDA-MB-231 confluent cells seeded in 24 well plates. The medium was replaced by fresh medium containing 5 µM docetaxel in the absence of PAH, or the presence of 5 µM PAH. Two pictures were taken at two different points of each well immediately after wound, and 3 days after wound with 5X objective. The width of the wound was measured using ImageJ software and reported as final wound width/initial wound width in percentage. Data presented are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Migration, Membrane, Staining, Microscopy, Wound Healing Assay, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 contributes to the efflux of doxorubicin from TNBC cells. A. PTCH1 protein expression (left panel) and intracellular doxorubicin (right panel) were analyzed 16 h after transfection of MDA-MB-231 cells with PTCH1-siRNA or negative-control-siRNA. PTCH1 and GAPDH western blot signals were quantified using ImageJ software (left panel). For doxorubicin accumulation measurements (right panel), MDA-MB-231 cells were grown on slides and transfected with PTCH1-siRNA or negative-control-siRNA. After incubation with 10 µM doxorubicin, 3 coverslips were fixed for doxorubicin loading control; the other coverslips were incubated with efflux buffer and fixed. B. Docetaxel inhibits the accumulation of doxorubicin in MDA-MB-231 cells. Cells on coverslip were incubated with 10 µM doxorubicin or 10 µM doxorubicin and 50 µM docetaxel. Doxorubicin fluorescence images were acquired by epifluorescence microscopy using a 40X objective, and doxorubicin fluorescence was quantified using ImageJ software for about 100 cells per condition per experiment. Histograms are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transfection, Negative Control, Western Blot, Software, Incubation, Control, Fluorescence, Epifluorescence Microscopy

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the proliferation of T24 cells. (A) RT-qPCR analysis of si-ARAF transfection efficiency. (B and C) Western blot analysis of si-ARAF transfection efficiency. (D) EdU fluorescence assay showing proliferating cells. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Quantitative RT-PCR, Transfection, Western Blot, Fluorescence

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on apoptosis of T24 cells. (A and B) Flow cytometry analysis of apoptosis levels. (C and D) Protein expression of cleaved-caspase 3 and total caspase 3. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. **p<0.01, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Flow Cytometry, Expressing

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the metastatic capacity of T24 cells. (A and B) Quantification of migrating cells. (A and C) Quantification of invading cells. (D and E) Expression of EMT-related proteins in T24 cells. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Expressing

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Effect of ARAF on the p38 MAPK pathway in T24 cells. (A) The level of p-p38, p38, p-ERK, ERK, p-MEK, MEK in T24 cells was determined by western blot assay. (B) p-p38/p38 ratio. (C) p-ERK/ERK ratio. (D) p-MEK/MEK ratio. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using one-way ANOVA. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Western Blot

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin activated p38 MAPK pathway in si-ARAF transfected T24 cells. (A) The level of p-p38, p38, p-ERK, ERK, p-MEK, MEK in T24 cells was determined by western blot assay. (B) p-p38/p38 ratio. (C) p-ERK/ERK ratio. (D) p-MEK/MEK ratio. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Transfection, Western Blot

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin reversed the effect of ARAF on T24 cell proliferation and apoptosis. (A) EdU fluorescence for cell proliferation capacity. (B) The apoptotic rate of T24 cells. (C and D) The protein level of Cleaved-caspase3 and Caspase3. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques: Fluorescence

Journal: Open Medicine

Article Title: ARAF regulates malignant progression of bladder cancer through the p38MAPK pathway

doi: 10.1515/med-2026-1422

Figure Lengend Snippet: Anisomycin reversed the effect of ARAF on T24 cell metastasis and EMT. (A–C) The migratory and invasive ability of T24 cells. (D–F) The protein level of E-cadherin and N-cadherin. Data are presented as mean ± SD from three independent biological replicates. Statistical analysis was performed using Student’s t-test. **p<0.01, ****p<0.0001.

Article Snippet: The human ureter epithelial immortalized cell line SV-HUC-1 (CL-0222) and human bladder cancer cell lines T24 (CL-0227) and J82 (CL-0125) were purchased from Wuhan Procell Biological Company.

Techniques:

Journal: Breast Cancer : Targets and Therapy

Article Title: The Regulatory Mechanism and Value in Early Diagnosis and Prognosis Assessment of miR-4448 in Breast Cancer

doi: 10.2147/BCTT.S592003

Figure Lengend Snippet: The effect of miR-4448 expression level on MDA-MB-231 and MCF-7 cells. ( A ) The effect of miR-4448 inhibitor/mimic. ( B ) The effect of miR-4448 inhibitor/mimic on cell migration. ( C ) The effect of miR-4448 inhibitor/mimic on cell invasion. The effect of miR-4448 inhibitor/mimic on cell proliferation in MDA-MB-231 cells ( D ) and MCF-7 cells ( E ). * P < 0.05, ** P < 0.01, *** P < 0.001; # P < 0.05, ## P < 0.01, ### P < 0.001.

Article Snippet: The human breast cancer cell line MDA-MB-231 was obtained from Procell (Wuhan, China) and cultured in Leibovitz’s L-15 medium containing 10% FBS and 1% PS.

Techniques: Expressing, Migration

Journal: Breast Cancer : Targets and Therapy

Article Title: The Regulatory Mechanism and Value in Early Diagnosis and Prognosis Assessment of miR-4448 in Breast Cancer

doi: 10.2147/BCTT.S592003

Figure Lengend Snippet: MiR-4448 and its target gene TPK1 . ( A ) TargetScan Database predicted the sequence of miR-4448 and TPK1 . ( B and C ) Dual-luciferase reporter gene in MDA-MB-231 and MCF-7 cells. ( D ) The expression level of TPK1 in the serum. ( E ) Pearson analysis between miR-4448 and TPK1 . ** P < 0.01, *** P < 0.001; ## P < 0.01.

Article Snippet: The human breast cancer cell line MDA-MB-231 was obtained from Procell (Wuhan, China) and cultured in Leibovitz’s L-15 medium containing 10% FBS and 1% PS.

Techniques: Sequencing, Luciferase, Expressing

Journal: Breast Cancer : Targets and Therapy

Article Title: The Regulatory Mechanism and Value in Early Diagnosis and Prognosis Assessment of miR-4448 in Breast Cancer

doi: 10.2147/BCTT.S592003

Figure Lengend Snippet: The effect of TPK1 expression level on MDA-MB-231 and MCF-7 cells. ( A ) The effect of si- TPK1 . ( B ) The effect of si- TPK1 on cell migration. ( C ) The effect of si- TPK1 on cell invasion. The effect of si- TPK1 on cell proliferation in MDA-MB-231 cells ( D ) and MCF-7 cells ( E ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human breast cancer cell line MDA-MB-231 was obtained from Procell (Wuhan, China) and cultured in Leibovitz’s L-15 medium containing 10% FBS and 1% PS.

Techniques: Expressing, Migration