Journal: Oncology Reports

Article Title: FGD5-AS1 is an oncogenic lncRNA in pancreatic cancer and regulates the Wnt/β-catenin signaling pathway via miR-577

doi: 10.3892/or.2021.8232

Figure Lengend Snippet: FGD5-AS1 directly targets miR-577. (A) Bioinformatics analysis was carried out to predict the binding sites between miR-577 and FGD5-AS1, and FGD5-AS1-WT and FGD5-AS1-MUT luciferase reporter gene vectors were constructed. (B) A dual-luciferase reporter assay was utilized to determine the binding site between FGD5-AS1 and miR-577. (C) Pearson's correlation analysis was employed to analyze the correlation between FGD5-AS1 expression and miR-577 expression in pancreatic cancer tissues. (D) An RNA pull-down assay was employed to validate the direct interaction between miR-577 and FGD5-AS1. (E) A RIP assay was carried out to confirm that the complex containing miR-577 and FGD5-AS1 in SW1990 cells was immunoprecipitated by anti-Ago2. All of the experiments were performed in triplicate. ***P<0.001. FGD5-AS1, FGD5 antisense RNA 1; miR, microRNA; WT, wild-type; MUT, mutant; RIP, RNA immunoprecipitation.

Article Snippet: Control siRNA (si-NC), FGD5-AS1 siRNA (si-FGD5-AS1), miR-577 mimics (miR-577), miR-577 inhibitors (miR-577 in), microRNA mimics control (miR-con), and microRNA inhibitor control (miR-con in) were obtained from Shanghai GenePharma Co., Ltd.

Techniques: Binding Assay, Luciferase, Construct, Reporter Assay, Expressing, Pull Down Assay, Immunoprecipitation, Mutagenesis

Journal: Oncology Reports

Article Title: FGD5-AS1 is an oncogenic lncRNA in pancreatic cancer and regulates the Wnt/β-catenin signaling pathway via miR-577

doi: 10.3892/or.2021.8232

Figure Lengend Snippet: Inhibition of miR-577 facilitates pancreatic cancer cell proliferation, migration and invasion. (A) RT-qPCR was utilized to detect miR-577 expression in the pancreatic cancer tissue and adjacent tissue samples (n=37). (B) RT-qPCR was employed to detect miR-577 expression in pancreatic cancer cell lines (SW1990, PANC-1, CAPAN-1 and BXPC-3) and normal pancreatic ductal epithelial cells (HPDE6-C7). (C) SW1990 cells were transfected with miR-con inhibitors or miR-577 inhibitors to construct the knockdown expression model of miR-577, and the transfection efficiency was detected by RT-qPCR. (D) An MTT assay was employed to detect the effect of miR-577 inhibition on SW1990 cell proliferation. (E) Transwell assays were utilized to detect the effects of miR-577 inhibition on SW1990 cell migration and invasion. Scale bar, 50 µm. All of the experiments were performed in triplicate. **P<0.01 and ***P<0.001. miR, microRNA RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Control siRNA (si-NC), FGD5-AS1 siRNA (si-FGD5-AS1), miR-577 mimics (miR-577), miR-577 inhibitors (miR-577 in), microRNA mimics control (miR-con), and microRNA inhibitor control (miR-con in) were obtained from Shanghai GenePharma Co., Ltd.

Techniques: Inhibition, Migration, Quantitative RT-PCR, Expressing, Transfection, Construct, MTT Assay, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: FGD5-AS1 is an oncogenic lncRNA in pancreatic cancer and regulates the Wnt/β-catenin signaling pathway via miR-577

doi: 10.3892/or.2021.8232

Figure Lengend Snippet: miR-577 targets LRP6 and β-catenin. (A) A dual-luciferase reporter assay was utilized to validate the binding sites between miR-577 and LRP6 3′UTR. (B) A dual-luciferase reporter assay was utilized to validate the binding sites between miR-577 and β-catenin 3′UTR. (C) Reverse transcription-quantitative PCR was employed to detect LRP6 and β-catenin mRNA expression levels in SW1990 cells transfected with miR-577 mimics. (D) A TOPFlash luciferase reporter assay was performed to detect the effects of miR-577 on the activity of the Wnt/β-catenin signaling. (E and F) Pearson's correlation analysis was utilized for the correlations between (E) FGD5-AS1 and LRP6 and between (F) FGD5-AS1 and β-catenin in pancreatic cancer tissues. (G and H) Pearson's correlation analysis was utilized for the correlations between (G) miR-577 and LRP6 and between (H) miR-577 and β-catenin in pancreatic cancer tissues. All of the experiments were performed in triplicate. **P<0.01 and ***P<0.001. LRP6, low-density lipoprotein receptor-related protein 6; miR, microRNA; FGD5-AS1, FGD5 antisense RNA 1; UTR, untranslated region.

Article Snippet: Control siRNA (si-NC), FGD5-AS1 siRNA (si-FGD5-AS1), miR-577 mimics (miR-577), miR-577 inhibitors (miR-577 in), microRNA mimics control (miR-con), and microRNA inhibitor control (miR-con in) were obtained from Shanghai GenePharma Co., Ltd.

Techniques: Luciferase, Reporter Assay, Binding Assay, Real-time Polymerase Chain Reaction, Expressing, Transfection, Activity Assay

Journal: Oncology Reports

Article Title: FGD5-AS1 is an oncogenic lncRNA in pancreatic cancer and regulates the Wnt/β-catenin signaling pathway via miR-577

doi: 10.3892/or.2021.8232

Figure Lengend Snippet: Inhibition of miR-577 partially reverses the biological behaviors of pancreatic cancer cells induced by FGD5-AS1 knockdown. (A) miR-577 inhibitors were transfected into SW1990 cells with FGD5-AS1 knockdown, and miR-577 expression in the transfected cells was detected by reverse transcription-quantitative PCR. (B) An MTT assay was utilized to detect the proliferation of SW1990 cells following transfection. (C) Transwell assays were utilized to detect the migration and invasion of SW1990 cells following transfection. Scale bar, 50 µm. (D) Western blotting was utilized to detect the protein expression of Axin2, cyclin D1 and c-Myc in SW1990 cells following transfection. All of the experiments were performed in triplicate. **P<0.01 and ***P<0.001. miR, microRNA; FGD5-AS1, FGD5 antisense RNA 1; si-, small interfering.

Article Snippet: Control siRNA (si-NC), FGD5-AS1 siRNA (si-FGD5-AS1), miR-577 mimics (miR-577), miR-577 inhibitors (miR-577 in), microRNA mimics control (miR-con), and microRNA inhibitor control (miR-con in) were obtained from Shanghai GenePharma Co., Ltd.

Techniques: Inhibition, Transfection, Expressing, Real-time Polymerase Chain Reaction, MTT Assay, Migration, Western Blot

Journal: Oncology Reports

Article Title: FGD5-AS1 is an oncogenic lncRNA in pancreatic cancer and regulates the Wnt/β-catenin signaling pathway via miR-577

doi: 10.3892/or.2021.8232

Figure Lengend Snippet: Graphical abstract. Long non-coding RNA FGD5-AS1 expression is markedly elevated in pancreatic cancer tissues and cells, and it could upregulate the expression of LPR6 and β-catenin by suppressing miR-577 expression via a ceRNA mechanism. Therefore, FGD5-AS1 modulated proliferation, migration and invasion of pancreatic cancer cells through miR-577/Wnt signaling axis. Lnc, long non-coding; miR, microRNA; ce, competing endogenous.

Article Snippet: Control siRNA (si-NC), FGD5-AS1 siRNA (si-FGD5-AS1), miR-577 mimics (miR-577), miR-577 inhibitors (miR-577 in), microRNA mimics control (miR-con), and microRNA inhibitor control (miR-con in) were obtained from Shanghai GenePharma Co., Ltd.

Techniques: Expressing, Migration

Journal: Journal of Breast Cancer

Article Title: LncRNA Taurine-Upregulated Gene 1 Promotes Cell Proliferation by Inhibiting MicroRNA-9 in MCF-7 Cells

doi: 10.4048/jbc.2016.19.4.349

Figure Lengend Snippet: si-NC=control si-RNA. * p <0.05 vs. si-NC; † p <0.05 vs. si- TUG1 ; ‡ p <0.05 vs. miR-9 inhibitor.

Article Snippet: Three small interfering RNAs (si-RNAs) specifically targeting TUG1 (si- TUG1 , si- TUG1 2, and si- TUG1 3) and the control si-RNA (si-NC) were purchased from Ribobio (Guangzhou, China).

Techniques: Control

Journal: Journal of Breast Cancer

Article Title: LncRNA Taurine-Upregulated Gene 1 Promotes Cell Proliferation by Inhibiting MicroRNA-9 in MCF-7 Cells

doi: 10.4048/jbc.2016.19.4.349

Figure Lengend Snippet: si-NC=control si-RNA; GRPDH=glyceraldehyde-3-phosphate dehydrogenase. * p <0.05 vs. si-NC; † p <0.05 vs. si- TUG1 .

Article Snippet: Three small interfering RNAs (si-RNAs) specifically targeting TUG1 (si- TUG1 , si- TUG1 2, and si- TUG1 3) and the control si-RNA (si-NC) were purchased from Ribobio (Guangzhou, China).

Techniques: Control

Journal: Stem Cell Research & Therapy

Article Title: Human liver organoids generated with single donor-derived multiple cells rescue mice from acute liver failure

doi: 10.1186/s13287-017-0749-1

Figure Lengend Snippet: Single donor-derived UC-ECs and UC-MCs promote spontaneous LO formation and differentiation. a Macroscopic view of spontaneously generated macro-LOs in coculture with human iPSC-endoderm, control ECs, and BM-MCs (con-EC/MC) or single-donor UC-derived ECs and MCs (donor UC-EC/MC, two donors) at 0 and 24 h. Dotted line represents the organoid area. Blue arrowhead, macro-LOs. Scale bar, 2 mm. b Percent areas of macro-LOs during spontaneous formation from 0 to 72 h. c Morphology of ΔEC/MC LOs (deletion of ECs and MCs in LOs), con-EC/MC LOs (containing control ECs and MCs in LOs), and donor UC-EC/MC LOs (containing donor UC-ECs and UC-MCs in LOs) on days 1 and 15. Scale bar, 100 μm. d ALB production capacities of ΔEC/MC LOs ( n = 4), con-EC/MC LOs ( n = 6), and donor UC-EC/MC LOs (four donors, n = 3 each) on day 15. e ALB , CYP3A4 , G6PC , and TDO2 expression in ΔEC/MC LOs (green, n = 4), con-EC/MC LOs (blue, n = 6), and donor UC-EC/MC LOs (red, four donors, n = 12) on day 15, as determined by qPCR. * p < 0.05, ** p < 0.01, *** p < 0.001. ALB albumin, ns not significant, Con control, EC endothelial cell, LO liver organoid, MC mesenchymal cell, UC umbilical cord, CYP cytochrome P450, G6PC glucose-6-phosphatase catalytic subunit, TDO2 tryptophan 2,3-dioxygenase

Article Snippet: ECs and BM-derived mesenchymal stem cells were obtained from Lonza as control ECs (con-ECs) and MCs (con-MCs) and maintained in EGM and MSCGM, respectively.

Techniques: Derivative Assay, Generated, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Human liver organoids generated with single donor-derived multiple cells rescue mice from acute liver failure

doi: 10.1186/s13287-017-0749-1

Figure Lengend Snippet: Single donor cell-derived LOs generated from single donor-derived hiPSC-endoderm, ECs, and MCs. a Schematic representation of the protocol for LO generation and differentiation from single donor-derived hiPSC-endoderm, ECs, and MCs. b Morphology of single donor cell-derived LOs (SDC-LOs) on days 1 and 15. Scale bars, 100 μm. c Immunocytochemical detection of ALB (red) and A1AT (green) expression in differentiated SDC-LOs on day 15. Scale bars, 100 μm. d Expression of common hepatic genes ( ALB , G6PC , RBP4 , TAT , and TDO2 ), urea/ammonia metabolism-related genes ( CPS1 and OTC ), and cytochrome P450-encoding genes ( CYP2C9 , CYP2C19 , CYP3A4 , CYP3A5 , and CYP3A7 ) in EC-iPSC-HLCs (blue, n = 6) and differentiated SDC-LOs (red, n = 6), determined by qPCR. e ALB production in EC-iPSC-HLCs ( n = 6), differentiated SDC-LOs ( n = 6), and primary human hepatocytes (PHHs, n = 4); data normalized to 1 million cells. f Urea production in EC-iPSC-HLCs ( n = 3), differentiated SDC-LOs ( n = 3), and PHHs ( n = 4); data normalized to 1 million cells. g Quantification of CYP3A4 activity in EC-iPSC-HLCs ( n = 4) and SDC-LOs ( n = 4) with or without rifampicin (25 μM) induction; data normalized to 1 million cells. *** p < 0.001. ns not significant, ALB albumin, EC endothelial cell, HGF hepatocyte growth factor, hiPSC human induced pluripotent stem cell, HLC hepatic-like cell, LO liver organoid, MC mesenchymal cell, PHH primary human hepatocyte, UC umbilical cord, CYP cytochrome P450, G6PC glucose-6-phosphatase catalytic subunit, TDO2 tryptophan 2,3-dioxygenase

Article Snippet: ECs and BM-derived mesenchymal stem cells were obtained from Lonza as control ECs (con-ECs) and MCs (con-MCs) and maintained in EGM and MSCGM, respectively.

Techniques: Derivative Assay, Generated, Expressing, Activity Assay