human urinary bladder transitional cell carcinoma tcc cell lines j82  (ATCC)

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