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: 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

Journal: International Journal of Molecular Sciences

Article Title: BMP9 Promotes the Proliferation and Migration of Bladder Cancer Cells through Up-Regulating lncRNA UCA1

doi: 10.3390/ijms19041116

Figure Lengend Snippet: The validation of recombinant adenovirus BMP9 and siBMP9. ( A ) The expression of BMP9 in normal bladder mucosa ( n = 126), superficial bladder cancer ( n = 68), and infiltrating bladder cancer ( n = 62) in the Lee Bladder database. p = 0.007; ( B ) The expression levels of BMP9 in T24 and BIU-87 cells were detected by western blot; ( C ) The BMP9 was up-regulated in BIU-87 cells after being transfected with AdBMP9 compared to the control group; ( D ) The BMP9 was down-regulated in T24 cells after being transfected with AdsiBMP9 compared to the control group. Data are shown as mean ± SD. ** p < 0.01.

Article Snippet: Human bladder cancer BIU-87 and T24 cells were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: Biomarker Discovery, Recombinant, Expressing, Western Blot, Transfection, Control

Journal: International Journal of Molecular Sciences

Article Title: BMP9 Promotes the Proliferation and Migration of Bladder Cancer Cells through Up-Regulating lncRNA UCA1

doi: 10.3390/ijms19041116

Figure Lengend Snippet: BMP9 up-regulated the expression of lncRNA UCA1 in bladder cancer cells. ( A ) Five common lncRNA were screened in BIU-87 cells after transfected with AdBMP9 by RT-PCR; ( B ) The expression of lncRNA UCA1 were verified in BIU-87 cells after transfected with AdBMP9 by RT-PCR; ( C ) The expression of lncRNA UCA1 were tested in T24 cells after being transfected with AdsiBMP9 by RT-PCR; ( D ) The inhibitory effect of siUCA1 were analyzed by RT-PCR in BIU-87 cells after being co-transfected with AdBMP9 and siUCA1. Data are shown as mean ± SD. ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, vs. control groups.

Article Snippet: Human bladder cancer BIU-87 and T24 cells were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: Gene expression of GDF 15 in human bladder cells . The mRNA levels of GDF15 (A) and α-SMA (B) in bladder smooth muscle cells (HBdSMC), fibroblast cells (HBdSF), normal epithelial cells (HBdEC), and carcinoma cell lines (RT-4, HT1376, T24, and TSGH-8301) were determined by RT-qPCR assays. Data are expressed as the mean ratio of mRNA in relation to HJBdEC cells (n = 3). (C) The protein levels of α-SMA, UPK2, and β-actin in bladder cells as indicated were determined by immunoblot assays. (D) GDF15 secretions from bladder cells as determined by ELISA (n = 4).

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: Effects of CAPE on GDF15 expression and proliferation of bladder carcinoma cells . (A) HT1376 and T24 cells were treated with various concentrations of CAPE as indicated for 24 h, then lysed, and the mRNA levels of GDF15 and β-actin were determined by RT-qPCR assays. Data are expressed as the mean ratio of mRNA relative to the control group (n = 3). (B) GDF15 protein levels in conditional media were determined by ELISA (n = 4). (C) T24 cells were treated with various concentrations of CAPE as indicated for 24 h and protein levels of GDF15 were determined by immunoblots. (D) The reporter activity of the GDF15 reporter vector in HT1376 cells treated with various dosages of CAPE for 24 h. Data are expressed as the mean percentage of luciferase activity relative to the mock-transfected group (n = 6). The mRNA (E) and protein (F) levels of target genes as indicated were determined by RT-qPCR and immunoblots. Data are expressed as the mean ratio of mRNA relative to the control group (n = 3). (G) Cell proliferation of T24 cells was determined by EdU assays after CAPE (30 μM) treatment for 24 h. Data are expressed as the mean percentage of Edu positive cells relative to the control group (DMSO-treated, n = 4). ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Expressing, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay, Plasmid Preparation, Luciferase, Transfection

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: Downregulation of cell proliferation and invasion by CAPE is GDF15-dependent in bladder carcinoma HT1376 cells . (A) HT_shCOL and HT_shGDF15 cells were treated with (+) or without (−) CAPE (30 μM) for 24 h. The protein levels of target genes, as indicated, were determined via immunoblotting. (B) HT_shCOL and HT_shGDF15 cells were treated with (+) or without (−) CAPE (30 μM) for 24 h and GDF15 secretions in the supernatants of culture media were determined by ELISA (n = 4). (C) HT_shCOL and HT_shGDF15 cells were treated with (+) or without (−) CAPE (30 μM) for 24 h, then the relative mRNA levels of GDF15, NDRG1, and maspin were determined by RT-qPCR assays (n = 3). (D) Cell proliferation of HT_shCOL and HT_shGDF15 cells after treated with CAPE (30 μM) for 48 h was determined by EdU assays (n = 4). (E) The quantitative result of EdU assays. (F) HT_shCOL and HT_shGDF15 cells were treated with 30 μM CAPE for 48 h and cell proliferation was determined by the CyQuant assay. The data were presented as the mean percentage compared with the control group (DMSO-treated, n = 8). (G) HT_shCOL and HT_shGDF15 cells were treated with CAPE (30 μM) for 24 h and the cell invasive ability was measured by the Matrigel invasion assay after 24 h of incubation (H) The quantitative result of invasion assays. The data were presented as the mean percentage compared with the control group (DMSO-treated, n = 3). ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, CyQUANT Assay, Control, Invasion Assay, Incubation

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: CAPE induces the phosphorylation of ERK, JNK, and p38 to modulate the expressions of GDF15, NDRG1, and maspin in bladder carcinoma HT1376 cells . (A) HT1376 cells were treated with various concentrations of CAPE as indicated for 24 h and the mRNA levels of target genes as indicated were determined by RT-qPCR assays. (B) Protein levels of target genes, as indicated, were determined by immunoblot assays after treatment with various concentrations of CAPE as indicated for 16 h in HT1376 cells. The quantitative data were expressed as the intensity of protein bands of the target genes/β-actin relative to the control solvent-treated group (n = 3). The expressions of ERK, p-ERK (C, top), JNK, p-JNK (D, top), p38, and p-p38 (E, top) were determined by the immunoblots after 20 min of 30 μM CAPE treatment with (+) or without (−) pretreatment of the indicated MAPK inhibitors for 1 h in HT1376 cells. The protein level of GDF15, NDRG1, maspin, and β-actin of HT1376 cells after CAPE treatment with (+) or without (−) pretreatment with PD0325901 (C, bottom), SP600125 (D, bottom), or SB202190 (E, bottom). The quantitative data were expressed as the intensity of protein bands of the p-target gene/target gene or target genes/β-actin relative to the control solvent-treated group (n = 3). ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Phospho-proteomics, Quantitative RT-PCR, Western Blot, Control, Solvent

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: CAPE inhibits tumor growth of HT1376 cells in xenograft mice model . The athymic male nude mice were subcutaneously injected with HT1376 cells and when tumor volumes reached around 75 mm 3 (day 13), they received vehicle (0.1% DMSO in PBS; n = 6) or CAPE (10 mg/kg; n = 6) injected once intraperitoneally per day, 5 days/week. (A) The mice were sacrificed and the tumors were collected and photographed. (B) Tumor volumes were measured in vehicle-treated (●) or CAPE-treated (○) groups. (C) The tumor weights were presented as mean tumor weight in grams. (D) The average body weight of mice was measured during the experimental period. (E) Whole cell lysates of randomly selected tumor samples (n = 3) from the vehicle or CAPE-treated groups were subjected to immunoblotting (F) and RT-qPCR (G) assays. ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Injection, Western Blot, Quantitative RT-PCR

Journal: Biomedical Journal

Article Title: Caffeic acid phenethyl ester inhibits the growth of bladder carcinoma cells by upregulating growth differentiation factor 15

doi: 10.1016/j.bj.2021.10.006

Figure Lengend Snippet: CAPE upregulates the activities of the AMPKα1/2 signaling pathway in bladder carcinoma cells . (A) HT1376 cells were treated with various concentrations of CAPE for 24 h, and the protein levels of the AMPKα1/2 and p-AMPKα1/2 were determined using immunoblot assays. (B) The expression of AMPKα1 and AMPKα2 was determined by RT-qPCR in HT_shCOL and HT_shAMPKα1/2 cells (n = 3). (C) The expression of AMPKα1/2 and β-actin was determined by immunoblotting in HT_shCOL and HT_shAMPKα1/2 cells. The protein level of AMPKα1/2 and β-actin was determined by immunoblot assays (D) and the mRNA level of GDF15 was determined by RT-qPCR assays (E; n = 3) in HT_shCOL and HT_shAMPKα1/2 cells after (+) or without (−) 30 μM CAPE treatment. (F) The proliferation of HT_shCOL and HT_shAMPKα1/2 cells was determined by the EdU assays after (+) or without (−) 30 μM CAPE treatment for 24 h (n = 4). ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The bladder transitional cell carcinoma cell lines, RT-4, HT1376, TSGH-8301, and T24 cells, were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured as described previously [ ].

Techniques: Western Blot, Expressing, Quantitative RT-PCR