Journal: Cancer research

Article Title: Development of novel aptamer-based targeted chemotherapy for bladder cancer

doi: 10.1158/0008-5472.CAN-21-2691

Figure Lengend Snippet: ( A ) Selection scheme of chemically modified RNA aptamers that can internalize into T24 human bladder cancer cells but not SV-huc-1 normal bladder urothelial cells. R: purine. Y: pyrimidine. ( B ) Predicted secondary structure of aptamer B1 using sfold. ( C ) Binding curve of aptamer B1 with T24 cells. An all-DNA version of B1 and a B1-derived scrambled 35-nt sequence were used as control. Representative data of three independent experiments are shown. ( D ) Confocal microscopy analysis showed that aptamer B1 internalization is cell type-specific. T24 and KU-7 are two bladder cancer cell lines, SV-huc-1 is a normal bladder urothelial cell line. Scale bars: 40 μm. Representative data of three independent experiments are shown. (E) Comparison of the binding curves of aptamer B1 with primary human tumor cells and normal urothelial cells generated from surgical specimens harvested from a NMIBC patient. Freshly collected bladder tumor tissue or normal para-tumor urothelium tissue was digested into single cells, incubated with Cy5.5-labeled aptamer B1 for 1 h and analyzed with flow cytometry. Raw data are shown in Supplementary Fig. 3B .

Article Snippet: Human bladder cancer cell lines T24 (CLS Cat# 300352/p619_T24, RRID: CVCL_0554) ( 27 ), TCCSUP (ATCC Cat# HTB-5, RRID: CVCL_1738), J82 (KCLB Cat# 30001, RRID: CVCL_0359), SW780 (ATCC Cat# CRL-2169, RRID: CVCL_1728), UM-UC-3 (KCB Cat# KCB 2014053YJ, RRID: CVCL_1783), RT4 (CLS Cat# 300326/p10281_RT-4, RRID: CVCL_0036) and Biu87 (KCB Cat# KCB 2006103YJ, RRID: CVCL_6881) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China) between 2018 and 2021.

Techniques: Selection, Modification, Binding Assay, Derivative Assay, Sequencing, Control, Confocal Microscopy, Comparison, Generated, Incubation, Labeling, Flow Cytometry

Journal: Cancer research

Article Title: Development of novel aptamer-based targeted chemotherapy for bladder cancer

doi: 10.1158/0008-5472.CAN-21-2691

Figure Lengend Snippet: ( A ) Cells were incubated with Cy5.5-labeled aptamer B1, lysosomes and nuclei were stained with LysoTracker Green and Hoechst 33342, respectively. Scale bar: 40 μm. ( B and C ) Cells were first treated with an endocytosis inhibitor, and then incubated with Cy5.5-labeled aptamer B1, followed by flow cytometry analysis. Chlorpromazine inhibits clathrin-mediated endocytosis. Amiloride inhibits pinocytosis. (D) SDS-PAGE analysis of proteins recovered from pull-down experiments using biotin-labeled aptamer B1. Asterisk indicates a protein band specific to aptamer B1 treatment. (E) Knockdown of AP2B1 with siRNA reduced aptamer B1 internalization into T24 cells. Representative data of three independent experiments are shown.

Article Snippet: Human bladder cancer cell lines T24 (CLS Cat# 300352/p619_T24, RRID: CVCL_0554) ( 27 ), TCCSUP (ATCC Cat# HTB-5, RRID: CVCL_1738), J82 (KCLB Cat# 30001, RRID: CVCL_0359), SW780 (ATCC Cat# CRL-2169, RRID: CVCL_1728), UM-UC-3 (KCB Cat# KCB 2014053YJ, RRID: CVCL_1783), RT4 (CLS Cat# 300326/p10281_RT-4, RRID: CVCL_0036) and Biu87 (KCB Cat# KCB 2006103YJ, RRID: CVCL_6881) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China) between 2018 and 2021.

Techniques: Incubation, Labeling, Staining, Flow Cytometry, SDS Page, Knockdown

Journal: Cancer research

Article Title: Development of novel aptamer-based targeted chemotherapy for bladder cancer

doi: 10.1158/0008-5472.CAN-21-2691

Figure Lengend Snippet: ( A ) Schematic of the self-assembled aptamer-tethered DNA nanotrain for cellular delivery of drugs. Aptamer directs drug-loaded nanotrain for selective internalization into bladder cancer cells. Chemotherapeutic drugs are unloaded inside the cell and induce cytotoxicity. Drug intrinsic fluorescence can be used to monitor its intercalation into DNA duplex and intracellular release. ( B ) Agarose gel analysis of nanotrain assembly. Lane1: M1+M2; Lane2: M1; Lane3: M2; Lane4: aptamer-tethered; Lane5: aptamer-tethered DNA nanotrain. ( C ) Flow cytometry analysis of nanotrain internalization into T24 and SV-huc-1 cells. ( D ) Confocal microscope images of nanotrain internalization into T24 and SV-huc-1 cells. Scale bars: 40 μm. Representative data of three independent experiments are shown.

Article Snippet: Human bladder cancer cell lines T24 (CLS Cat# 300352/p619_T24, RRID: CVCL_0554) ( 27 ), TCCSUP (ATCC Cat# HTB-5, RRID: CVCL_1738), J82 (KCLB Cat# 30001, RRID: CVCL_0359), SW780 (ATCC Cat# CRL-2169, RRID: CVCL_1728), UM-UC-3 (KCB Cat# KCB 2014053YJ, RRID: CVCL_1783), RT4 (CLS Cat# 300326/p10281_RT-4, RRID: CVCL_0036) and Biu87 (KCB Cat# KCB 2006103YJ, RRID: CVCL_6881) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China) between 2018 and 2021.

Techniques: Fluorescence, Agarose Gel Electrophoresis, Flow Cytometry, Microscopy

Journal: Cancer research

Article Title: Development of novel aptamer-based targeted chemotherapy for bladder cancer

doi: 10.1158/0008-5472.CAN-21-2691

Figure Lengend Snippet: ( A ) Characterization of the loading of epirubicin into the nanotrain. The intrinsic fluorescence of EPI decreased with increasing equivalents of nanotrain boxcar component strands, as it was quenched upon intercalation into DNA duplex. ( B ) Stability of the EPI-loaded nanotrain in PBS was evaluated by dialysis. Free EPI molecules diffused across the dialysis membrane and resulted in a high fluorescence intensity in the exterior chamber. Interaction of EPI with nanotrain (NT-EPI) restricted its diffusion. (C) Release of EPI in the presence of DNase. Intercalation of EPI into boxcar DNA duplex of NT resulted in fluorescence quenching. Treatment with DNase Ⅰ degraded dsDNA and restored the intrinsic fluorescence of EPI. ( D ) Confocal microscope images showing targeted delivery of NT-EPI into T24 and KU-7 bladder cancer cells. Cells were treated with free EPI or NT-EPI for 1 h. The nuclei were stained with DAPI. Scale bar: 40 μm. ( E and F ) Aptamer B1 NT-EPI exhibits similar cytotoxicity to that of free EPI against T24 (E) and KU-7 (F) bladder cancer cells. ( G ) Aptamer B1 NT-EPI was much less toxic to SV-huc-1 cells compared with free EPI. Cells were treated for 48 h and cell viability was measured by MTT assay. Data are mean ± SD. Representative data of three independent experiments shown.

Article Snippet: Human bladder cancer cell lines T24 (CLS Cat# 300352/p619_T24, RRID: CVCL_0554) ( 27 ), TCCSUP (ATCC Cat# HTB-5, RRID: CVCL_1738), J82 (KCLB Cat# 30001, RRID: CVCL_0359), SW780 (ATCC Cat# CRL-2169, RRID: CVCL_1728), UM-UC-3 (KCB Cat# KCB 2014053YJ, RRID: CVCL_1783), RT4 (CLS Cat# 300326/p10281_RT-4, RRID: CVCL_0036) and Biu87 (KCB Cat# KCB 2006103YJ, RRID: CVCL_6881) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China) between 2018 and 2021.

Techniques: Stable Transfection, Fluorescence, Membrane, Diffusion-based Assay, Microscopy, Staining, MTT Assay

Journal: Cancer research

Article Title: Development of novel aptamer-based targeted chemotherapy for bladder cancer

doi: 10.1158/0008-5472.CAN-21-2691

Figure Lengend Snippet: ( A ) Schematic of orthotopic xenograft model establishment and treatment regimen. Luciferase-labeled KU-7 human bladder cancer cells were implanted into the bladder of nude mice. Fourteen days later, mice with positive bladder fluorescence signal were randomly divided into three groups and treated with intravesical instillation of PBS, free EPI (0.4 mg/ml, 50 μl/instillation) or aptamer B1 NT-EPI (equal molar EPI molecules). ( B ) Luminescence imaging and ( C ) quantitative analysis of tumors in mouse bladder at the end of treatment. Data are mean ± SD (n = 9 mice/group). ** P < 0.01. ( D ) Representative H&E staining images of bladder tumor sections from each group. Sections of whole bladder was shown in the middle (scale bar: 500 μm), with magnification on both sides (scale bar: 100 μm). The NT-EPI group showed much less cystitis compared to free EPI, as indicated by reduced infiltration of inflammatory cells and interstitial edema, suggesting reduced damage to normal urothelium. Quantification of all nine mice in each group are summarized in Supplementary Fig. 6 .

Article Snippet: Human bladder cancer cell lines T24 (CLS Cat# 300352/p619_T24, RRID: CVCL_0554) ( 27 ), TCCSUP (ATCC Cat# HTB-5, RRID: CVCL_1738), J82 (KCLB Cat# 30001, RRID: CVCL_0359), SW780 (ATCC Cat# CRL-2169, RRID: CVCL_1728), UM-UC-3 (KCB Cat# KCB 2014053YJ, RRID: CVCL_1783), RT4 (CLS Cat# 300326/p10281_RT-4, RRID: CVCL_0036) and Biu87 (KCB Cat# KCB 2006103YJ, RRID: CVCL_6881) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China) between 2018 and 2021.

Techniques: Inhibition, In Vivo, Luciferase, Labeling, Fluorescence, Imaging, Staining

Journal: Mutation research

Article Title: A pilot study exploring time- and dose-dependent DNA damage and chromosomal instability caused by benzo[a]pyrene in two urothelial cell types.

doi: 10.1016/j.mrfmmm.2024.111855

Figure Lengend Snippet: Fig. 1. Cellular metabolic activity. Relative change in viability measured by the amount of formazan (MTT; (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) in (A) T24 and (B) PUBEC cells, (n = 2).

Article Snippet: T24 cells (ACC 376 purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany) were cultured in McCoy’s 5 A (c.c.pro) with 1% penicillin/streptomycin, 10% FCS, and 1% L-glutamine.

Techniques: Activity Assay

Journal: Mutation research

Article Title: A pilot study exploring time- and dose-dependent DNA damage and chromosomal instability caused by benzo[a]pyrene in two urothelial cell types.

doi: 10.1016/j.mrfmmm.2024.111855

Figure Lengend Snippet: Fig. 4. DNA damage. DNA damage assessed by comet assay and expressed as the percentage of DNA in tail for (A) T24 and (B) PUBEC cells. The results represent average tail intensities relative to control conditions (n = 1).".

Article Snippet: T24 cells (ACC 376 purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany) were cultured in McCoy’s 5 A (c.c.pro) with 1% penicillin/streptomycin, 10% FCS, and 1% L-glutamine.

Techniques: Single Cell Gel Electrophoresis, Control

Journal: Mutation research

Article Title: A pilot study exploring time- and dose-dependent DNA damage and chromosomal instability caused by benzo[a]pyrene in two urothelial cell types.

doi: 10.1016/j.mrfmmm.2024.111855

Figure Lengend Snippet: Fig. 3. Colony growth as a percentage of colony area in T24 and PUBEC cells (n = 1).

Article Snippet: T24 cells (ACC 376 purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany) were cultured in McCoy’s 5 A (c.c.pro) with 1% penicillin/streptomycin, 10% FCS, and 1% L-glutamine.

Techniques:

Journal: BMC Cancer

Article Title: Mechanisms for fibrate lipid-lowering drugs in enhancing bladder cancer immunotherapy by inhibiting CD276 expression

doi: 10.1186/s12885-025-14855-w

Figure Lengend Snippet: Fenofibrate (FNF) enhanced T cell-mediated antitumor immunity in bladder cancer. After T24 cells were pretreated with 0, 50, or 100 µM fenofibrate (FNF) for 24 h: A-B ELISA was used to detect the secretion levels of IFN-γ and TNF-α in the culture supernatants; C Pretreated T24 cells were co-cultured with activated T cells to evaluate the effect of FNF on T cell-mediated cytotoxicity against T24 cells; scale bar = 100 μm. ***, p < 0.001

Article Snippet: After incubation with 50 μM or 100 μM FNF for 24 h, cell culture supernatants were collected following centrifugation at 150 g for 15 min. IFN-γ (E-EL-H0108c) and TNF-α (E-EL-H0109c) concentrations in T24 cell supernatants were detected using ELISA kits purchased from Wuhan Elabscience Biotechnology Co., Ltd.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture