human bladder carcinoma cell line j82  (ATCC)

Journal: The FASEB Journal

Article Title: Intravesical Delivery of P21 mRNA –Loaded Lipid Nanoparticles as a Tumor Suppressor Replacement Therapy for Bladder Cancer

doi: 10.1096/fj.202600049R

Figure Lengend Snippet: Rationale for selecting CDKN1A /p21 as a therapeutic target in bladder cancer and validation of p21 mRNA expression. (A) Pan‐cancer analysis of CDKN1A alteration frequencies based on TCGA datasets. The analysis was performed using the TIMER3 web server. (B) CDKN1A mRNA expression in bladder cancer and normal bladder tissues, shown as log2(TPM + 1), where TPM denotes transcripts per million. Data were analyzed using the TIMER3 web server. (C) Representative p21 immunohistochemical staining images from a bladder cancer tissue microarray stratified by pathological stage (Tis, T1, T2, T3, and T4). Scale bars, 200 μm. (D) Quantification of p21 H‐scores in Tis, T1, T2, T3, and T4 lesions. Compared with Tis lesions, T1, T2, T3, and T4 tumors showed significantly lower p21 H‐scores. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01 (one‐way ANOVA). (E) Western blot analysis of basal and p21 mRNA‐induced p21 protein expression in T24 and J82 bladder cancer cells. Basal p21 protein levels were very low in both cell lines, whereas transfection of p21 mRNA led to robust p21 expression. For p21 blots, both short‐exposure (se) and long‐exposure (le) images are shown. β‐Actin serves as a loading control. (F) Immunofluorescence analysis of EGFP‐HA and p21‐HA expression in T24 and J82 cells using an anti‐HA antibody. Nuclei were stained with DAPI. The p21‐HA signal was predominantly localized to the nucleus, whereas EGFP showed diffuse cytoplasmic distribution. Scale bars, 20 μm.

Article Snippet: The human bladder carcinoma cell line J82 and the human embryonic kidney cell line HEK293T were purchased from the American Type Culture Collection (ATCC).

Techniques: Biomarker Discovery, Expressing, Immunohistochemical staining, Staining, Microarray, Western Blot, Transfection, Control, Immunofluorescence

Journal: International journal of molecular sciences

Article Title: Hispolon Methyl Ether, a Hispolon Analog, Suppresses the SRC/STAT3/Survivin Signaling Axis to Induce Cytotoxicity in Human Urinary Bladder Transitional Carcinoma Cell Lines.

doi: 10.3390/ijms24010138

Figure Lengend Snippet: Figure 1. HME’s cytotoxic effect on human bladder TCC cell lines. (A) HME induces stronger cytotoxicity than cisplatin against human bladder TCC cells. Human bladder TCC cell lines J82, T24, and TCCSUP were treated with HME or cisplatin (0~200 µM) for 48 h, followed by cell viability determination using MTS assay. (B) HME impairs the colony-forming capacity of human bladder TCC cells. The levels of clonogenicity of HME-treated J82, T24, and TCCSUP cells were determined as described in Section 4. ***: p < 0.001. (C) Higher cytotoxic effect of HME than hispolon on TCCSUP cells while comparable cytotoxicity of these two drugs against J82 and T24 cells. J82, T24, and TCCSUP were treated with HME or hispolon (0~200 µM) for 48 h, followed by cell viability determination using MTS assay.

Article Snippet: Human urinary bladder transitional cell carcinoma cell lines J82 (ATCC HTB-1TM) and TCCSUP (ATCC HTB-5TM) were both cultured in Eagle’s Minimum Essential Medium, while the T24 cell line (ATCC HTB-4TM) in McCoy’s 5a medium in accordance with the recommendation of the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: MTS Assay

Journal: International journal of molecular sciences

Article Title: Hispolon Methyl Ether, a Hispolon Analog, Suppresses the SRC/STAT3/Survivin Signaling Axis to Induce Cytotoxicity in Human Urinary Bladder Transitional Carcinoma Cell Lines.

doi: 10.3390/ijms24010138

Figure Lengend Snippet: Figure 2. HME’s proapoptotic effect on human bladder TCC cell lines. (A) Induction of PARP cleavage by HME. J82, T24, and TCCSUP cells were treated with HME (0, 30, 60 µM) for 24 h, followed by immunoblotting to evaluate the levels of PARP cleavage. The levels of glyceraldehyde- 3-phosphate dehydrogenase (GAPDH) were used as a control for equal loading. c-PARP: cleaved PARP. (B) Increase of annexin V-positive (apoptotic) cell population by HME. J82, T24, and TCCSUP cells were treated with HME (0, 30, 60 µM) for 24 h, followed by flow cytometry-based annexin V/propidium iodide (PI) dual staining as described in Section 4. ***: p < 0.001.

Article Snippet: Human urinary bladder transitional cell carcinoma cell lines J82 (ATCC HTB-1TM) and TCCSUP (ATCC HTB-5TM) were both cultured in Eagle’s Minimum Essential Medium, while the T24 cell line (ATCC HTB-4TM) in McCoy’s 5a medium in accordance with the recommendation of the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Western Blot, Control, Cytometry, Staining

Journal: International journal of molecular sciences

Article Title: Hispolon Methyl Ether, a Hispolon Analog, Suppresses the SRC/STAT3/Survivin Signaling Axis to Induce Cytotoxicity in Human Urinary Bladder Transitional Carcinoma Cell Lines.

doi: 10.3390/ijms24010138

Figure Lengend Snippet: Figure 5. HME inhibits SRC to suppress the STAT3/survivin signaling axis. (A) HME inhibits SRC activation in all human bladder TCC cell lines examined. J82, T24, and TCCSUP cells were treated with HME (0, 30, 60 µM) for 24 h, followed by immunoblotting for the levels of tyrosine 1007/1008- phosphorylated JAK2 (p-JAK2), total JAK2, tyrosine 416-phosphorylated SRC (p-SRC), total SRC, and GAPDH (loading control). (B) Ectopic expression of v-src, a dominant-active SRC, rescues both p-STAT3 and survivin levels from HME-mediated suppression but also attenuates HME-induced PARP cleavage. TCCSUP stable clones of HA-v-src and the respective vector control were treated with HME (0, 30, 60 µM) for 24 h, followed by immunoblotting to evaluate the levels of HA, p-STAT3, total STAT3, survivin, c-PARP, and GAPDH (loading control). (C) v-src overexpression antagonizes the proapoptotic and cytotoxic effect of HME. TCCSUP stable clones of HA-v-src and the respective vector control were treated with HME (0, 30, 60 µM) for 24 h, followed by annexin V/PI dual staining assay (left) and clonogenicity assay (right) to examine the HME-induced apoptosis and cytotoxicity, respectively. *: p < 0.05; ***: p < 0.001.

Article Snippet: Human urinary bladder transitional cell carcinoma cell lines J82 (ATCC HTB-1TM) and TCCSUP (ATCC HTB-5TM) were both cultured in Eagle’s Minimum Essential Medium, while the T24 cell line (ATCC HTB-4TM) in McCoy’s 5a medium in accordance with the recommendation of the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Activation Assay, Western Blot, Control, Expressing, Clone Assay, Plasmid Preparation, Over Expression, Staining

Journal: Cancers

Article Title: A Novel Cyclic Pentadepsipeptide, N -Methylsansalvamide, Suppresses Angiogenic Responses and Exhibits Antitumor Efficacy against Bladder Cancer

doi: 10.3390/cancers13020191

Figure Lengend Snippet: MSSV-mediated inhibition of proliferation of the human bladder cancer cell lines, 5637 and T24, was owing to G1-phase cell cycle arrest. ( A ) Both cell lines were treated with or without MSSV at the indicated concentrations for 24 h, followed by MTT assays for cell viability. ( B ) Cells were treated with indicated concentrations of MSSV for 24 h, and cell counting was performed via trypan blue staining. ( C ) Both cell lines were treated with MSSV for 24 h and analyzed for cell cycle distribution using fluorescence-activated cell sorting (FACS) histograms. The percentage of cells in each cell cycle phase is presented. ( D ) Cells were exposed to MSSV at the indicated concentrations for 24 h. The expression levels of cyclin D1, cyclin E, CDK2, CDK4, p21WAF1, p27KIP1, p53, and GAPDH were analyzed via immunoblotting. Bar graphs show the relative fold changes in proteins at different MSSV concentrations in comparison with the control. ( E ) The cell lysates were immunoprecipitated with antibodies recognizing CDK2 and CDK4, followed by immunoblotting with specific antibodies against p21WAF1, p27KIP1, CDK2, and CDK4. Graphs show the relative amount of immunoprecipitated proteins as fold changes in comparison with the control. For the bar graphs, values were presented as the mean ± SD of three independent experiments; * p < 0.05, compared with the control group. Uncropped Western Blot Images in .

Article Snippet: The human bladder carcinoma cell lines, 5637 and T24, were purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a 5% CO 2 humidified incubator.

Techniques: Inhibition, Cell Counting, Staining, Fluorescence, FACS, Expressing, Western Blot, Comparison, Control, Immunoprecipitation