Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: cDNA array analysis identified AKR1C1 as a potential target of avasimibe. (A) RBE cells were treated with avasimibe at the concentration of 20 µM for 24 and 48 hours and subjected to cDNA array analysis. Cluster of changed genes in cell proliferation was presented. (B) The level of AKR1C1 mRNA and protein was detected by RT-PCR and western blotting in RBE cells when treated with avasimibe (20µM) for 48 hours. (C) The expression of AKR1C1 and PCNA was detected by IHC on the resected xenografts. IHC, ×200. *P < 0.05, **P < 0.01. ‘long scores of AKR1C1’ means AKR1C1 staining score. IHC, immunohistochemistry.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: The oncogenic role of AKR1C1 in cholangiocarcinoma. (A) Representative images of negative staining of AKR1C1 in noncancerous tissues (N) and high expression of AKR1C1 in CCA (T). IHC,×200 for small pictures and×40 for large pictures. *P < 0.05. (B) Patients with AKR1C1 expression had a shorter time to recurrence (B1) and a worse overall survival (B2) than those without AKR1C1 expression. (C, D) QBC939 (C) and RBE (D) cells were transfected with AKR1C1-shRNA for 48 hours and the level of AKR1C1 mRNA was detected by RT-PCR. CCK8 assay was used to measure the cell viability of both cell lines. **P < 0.01, ****P < 0.0001. (E) QBC939 and RBE cells were treated with avasimibe with or without exogenous AKR1C1 plasmid for 48 hours and CCK8 was used to detect the changes of cell viability. IHC, immunohistochemistry; CCK8, Cell Counting Kit-8; Vec, vector.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Negative Staining, Expressing, Transfection, shRNA, Reverse Transcription Polymerase Chain Reaction, CCK-8 Assay, Plasmid Preparation, Immunohistochemistry, Cell Counting

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: Correlation between AKR1C1 expression and clinicopathological factors.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: Univariate and multivariate analysis of time to progression in 49 patients with hilar cholangiocarcinoma according to clinicopathologic factors and AKR1C1 overexpression.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Over Expression

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: Univariate and multivariate analysis of overall survival in 49 patients with hilar cholangiocarcinoma according to clinicopathological factors and AKR1C1 overexpression.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Over Expression

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: AKR1C1 is regulated by FoxM1 in cholangiocarcinoma. (A) RBE cells were treated with avasimibe (20 µM) for 24 and 48 hours and subjected to cDNA array analysis. Cluster of changed genes in regulation of transcription was presented. (B) The level of FoxM1 mRNA and protein was detected by RT-PCR and western blotting in RBE cells when treated with avasimibe (20µM) for 48 hours. (C) RBE cells were transfected with FoxM1-shRNA for 48 hours and the levels of FoxM1 and AKR1C1 mRNA and proteins were detected by RT-PCR and western blotting. (D) RBE cells were transfected with FoxM1 plasmid for 48 hours and the levels of FoxM1 and AKR1C1 mRNA and proteins were detected by RT-PCR and western blotting. *P < 0.05, ****P < 0.0001.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, shRNA, Plasmid Preparation

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: AKR1C1 is a direct transcriptional target of FoxM1. (A) Diagram shows the sequence and position of five putative FoxM1-binding elements in the AKR1C1 promoter. TSS, transcriptional start site; WT, wild type; Mut, mutant type. (B) Left panel, RBE cells were cotransfected with the AKR1C1 promoter reporter, pRL-TK, and pcDNA3.1-FoxM1 or pcDNA 3.1; right panel, RBE cells were cotransfected with the AKR1C1 promoter reporter, pRL-TK, and FoxM1-shRNA or shcontrol (50 nM). 36 hours after transfection, the cells were collected, and the relative AKR1C1 promoter activities were measured. The assay was repeated three times independently. ***P < 0.001. (C) Reporter plasmids harboring the wild-type AKR1C1 promoter or the corresponding mutant promoter in the FoxM1-binding sites were transfected into RBE cells, and the relative promoter activities were measured as above. (D) The chromatin immunoprecipitation (ChIP) assay results show the in vivo binding of FoxM1 to the AKR1C1 promoter. QBC939 cell lysis was immunoprecipitated using an anti-FoxM1 antibody or immunoglobulin G The resulting samples were subjected to RT-PCR using the site-specific primers.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Sequencing, Binding Assay, Mutagenesis, shRNA, Transfection, Chromatin Immunoprecipitation, In Vivo, Lysis, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Oncology

Article Title: Avasimibe Dampens Cholangiocarcinoma Progression by Inhibiting FoxM1-AKR1C1 Signaling

doi: 10.3389/fonc.2021.677678

Figure Lengend Snippet: Avasimibe inhibits cholangiocarcinoma cells proliferation via targeting AKR1C1 and FoxM1. (A, B) RBE cells (A) and QBC939 cells (B) were treated with avasimibe or DMSO, then transfected with FoxM1 or control vector, along with the transfection with AKR1C1 shRNA or shNT. 48 h after transfection, cell viability was analyzed by CCK8 assay. Data are from three independent assays. *P < 0.05, **P < 0.01, ***P < 0.001. (C) Left panel, the expression of FoxM1 and AKR1C1 was detected by IHC on the resected xenografts. IHC, ×400. Right panel, diagram showing the different expression of FoxM1 and AKR1C1 in these samples when treated with avasimibe. (D) The representative images of FoxM1 and AKR1C1 expressions and their correlations determined by Spearman’s correlation test. r, Spearman correlation coefficient; IHC, ×40 or ×200.

Article Snippet: FoxM1 (sc-500, Santa Cruz Biotechnology), PCNA (sc-500, Maixin-Bio, Fuzhou, China), and AKR1C1 (PB1091, Boster Biological Technology, Wuhan, China) antibodies were used for IHC analysis.

Techniques: Transfection, Control, Plasmid Preparation, shRNA, CCK-8 Assay, Expressing