Journal: BMB Reports

Article Title: Drug resistance of bladder cancer cells through activation of ABCG2 by FOXM1

doi: 10.5483/BMBRep.2018.51.2.222

Figure Lengend Snippet: Decreased FOXM1 reduces drug efflux and cell viability in BC cells. The cell viability was measured by MTT (A) and clonogenic assays (B). Cells were transfected with siRNA (scRNA and siFOXM1) or overexpression vector (pcDNA and pFOXM1) for 24 h and then exposed to 0.5 μM DOX for indicated time (C). Drug efflux activity was measured using a SP assay. KU7 cells were transfected with siRNA for 24 h. Cells were stained with Hoechst 33342 dye (1 mg/ml) in the presence or absence of the drug efflux inhibitor verapamil (100 μg/ml) and analyzed by FACS. (D) Effect of FOXM1 on apoptosis in BC cells. Cells were transfected with scRNA or siFOXM1 and then exposed to 0.5 μM DOX for 24 h. The apoptosis rate was measured using a Muse Annexin V kit.

Article Snippet: The plasmid DNA used was pGL3-basic, pGL3-basic-ABCG2 promoter, pcDNA6, pFOXM1 and pRL Renilla luciferase control reporter vector (Promega).

Techniques: Transfection, Over Expression, Plasmid Preparation, Activity Assay, Staining

Journal: BMB Reports

Article Title: Drug resistance of bladder cancer cells through activation of ABCG2 by FOXM1

doi: 10.5483/BMBRep.2018.51.2.222

Figure Lengend Snippet: FOXM1 regulates ABCG2 expression in BC cells. pcDNA and pFOXM1 were transfected into KU7 (A) and 5637 cells (B). scRNA and siFOXM1 were transfected into KU7 (C) and 5637 cells (D). After 24 h, mRNA and protein levels were analyzed using qRT-PCR (left panel) and Western blotting (right panel).

Article Snippet: The plasmid DNA used was pGL3-basic, pGL3-basic-ABCG2 promoter, pcDNA6, pFOXM1 and pRL Renilla luciferase control reporter vector (Promega).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: BMB Reports

Article Title: Drug resistance of bladder cancer cells through activation of ABCG2 by FOXM1

doi: 10.5483/BMBRep.2018.51.2.222

Figure Lengend Snippet: FOXM1 binds directly to the ABCG2 promoter to regulate transcription. (A) Schematic diagram of ABCG2 promoter vector (ABCG2P). The ABCG2 promoter (−2106/+22) was inserted into the pGL3 basic vector and the three putative FOXM1 binding sites (−1946/−1934, −1070/−1059 and −279/−268) are represented. The black bars on the promoter region indicate the positions of the primers (I: −2098/−1903, II: −1144/−1027, III: −326/−198 and non-target site (NTS): −1489/−1388) for qChIP amplification. (B) BC cells were transformed with pGL3-basic, ABCG2P or ABCG2P + pFOXM1, respectively. (C) BC cells were transformed with pGL3-basic, ABCG2P or ABCG2P + siFOXM1, respectively. transcriptional activity was measured by luciferase assay. (D) ChIP assay in the ABCG2 promoter. 5637 cells were transfected with pcDNA or pFOXM1 + V5 tag vector, Immunoprecipitation was performed using the rabbit IgG (control), FOXM1 and V5 antibody. The chromatin fragments were amplified using primers for the three putative FOXM1 binding sites (loci I, II, III) and NTS primers shown in (A).

Article Snippet: The plasmid DNA used was pGL3-basic, pGL3-basic-ABCG2 promoter, pcDNA6, pFOXM1 and pRL Renilla luciferase control reporter vector (Promega).

Techniques: Plasmid Preparation, Binding Assay, Amplification, Transformation Assay, Activity Assay, Luciferase, Transfection, Immunoprecipitation, Control

Journal: BMB Reports

Article Title: Drug resistance of bladder cancer cells through activation of ABCG2 by FOXM1

doi: 10.5483/BMBRep.2018.51.2.222

Figure Lengend Snippet: FOXM1 reduction was shown to decrease ABCG2 expression under DOX exposure. Two bladder cancer cells were transfected with siRNA (scRNA and siFOXM1) or overexpression vector (pcDNA and pFOXM1). After 24 h, transfected cells were exposed to 0.5 μM DOX for 24 h. mRNA (A) and protein (B, C) levels of FOXM1 and ABCG2 were analyzed by qPCR and western blotting methods.

Article Snippet: The plasmid DNA used was pGL3-basic, pGL3-basic-ABCG2 promoter, pcDNA6, pFOXM1 and pRL Renilla luciferase control reporter vector (Promega).

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Western Blot