Journal: Scientific Reports

Article Title: Linearized texture of three-dimensional extracellular matrix is mandatory for bladder cancer cell invasion

doi: 10.1038/srep36128

Figure Lengend Snippet: Texture of electrospun scaffolds made of PCL fibers deposited on drum collector ( a ) and grid collector ( b ) were investigated by scanning electron microscopy (SEM) and assessed for the ability to support binding and invasion of metastatic T24 cells. The degree of fibril organization (anisotropy index) was estimated on the SEM micrographs by using ImageJ. Presence of T24 cells on electrospun PCL was analyzed 7 days after seeding; histology evaluation was performed on sections obtained from a transverse cut of the scaffold. Metastatic T24 cells were seeded onto MIBC-derived ECM, and 4 days after seeding 3 μm paraffin sections from each sample were used, and hematoxylin-eosin stained sections were histologically evaluated ( c ) one experiment representative of 3 independently performed on MIBC-ECM obtained from 3 different tumors is shown. Cell viability of cells seeded for 7 days on the synthetic scaffold was compared to T24 cells plated on glass-slide (green and red spots show live and dead cells, respectively) ( d ) For panels a,b and d one experiment representative of 3 independently performed is shown.

Article Snippet: T24 cell line (CLS Cell Lines Service GmbH, Eppelheim, Germany) was seeded onto bladder tumor-derived ECM as previously reported .

Techniques: Electron Microscopy, Binding Assay, Derivative Assay, Staining

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