Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: AP000695.2 promotes TEAD1-mediated transcription of GLUT1 (A) Spearman correlation analysis in The Cancer Genome Atlas (TCGA) database showed that there is a positive correlation between AP000695.2 expression and TEAD1 mRNA expression (r = 0.2829). A significant positive correlation between the expression of TEAD1 mRNA and the mRNA expression of various glycolysis-related factors was observed. (B) Cellular experiments showed that the protein expression level of TEAD1 was increased after overexpression of AP000695.2, but decreased after the downregulation of AP000695.2. (C) In the GLUT1 Promoter, wt group, the expression of luciferase was significantly increased after overexpression of TEAD1. In the GLUT1 Promoter, mt group, the expression of luciferase was significantly increased after overexpression of TEAD1, and the expression level of luciferase in GLUT1 Promoter, wt group was significantly higher than that of GLUT1 Promoter, mt group after overexpression of TEAD1. MCS indicates negative control while TEAD1 indicates TEAD1 overexpression plasmid. (D) The expression of TEAD1 in TEAD1 overexpression (TEAD1) group was significantly higher than that in the NC group. (E) qPCR analysis showed that the mRNA expression level of GLUT1 in A549 and H1299 cell lines was increased after upregulating the expression of TEAD1. (F) Western blot analysis showed that the protein expression levels of GLUT1, HK2, PKM2 and LDHA in A549 and H1299 cell lines were increased after upregulating the expression of TEAD1. (G) Western blot analysis showed that the protein expression level of HIF-1α in A549 and H1299 cell lines was increased after upregulating the expression of TEAD1. * P<0.05, **P<0.01, ***P <0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Expressing, Over Expression, Luciferase, Negative Control, Plasmid Preparation, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: AP000695.2 promotes LUAD glycolysis, proliferation and migration via TEAD1 (A) The relative amount of FDG uptake of the L+sT group (transfected with AP000695.2 overexpression plasmid and TEAD1 siRNA) was lower than that of the L+s-NC (transfected with AP000695.2 overexpression plasmid and TEAD1 siRNA NC) group. The amount of FDG uptake in the s+T (transfected with AP000695.2 siRNA and TEAD1 overexpression plasmid) group was higher than that of the s+T-NC group (transfected with AP000695.2 siRNA and NC of TEAD1 overexpression plasmid). (B) The amount of lactic acid production in the L+sT group was lower than that in the L+s-NC group. The amount of lactic acid produced in the s+T group was higher than that in the s+T-NC group. (C) The level of glycolysis, glycolytic capacity and glycolytic reverse in the L+sT group were lower than those in the L+s-NC group. The level of glycolysis, glycolytic capacity and glycolytic reverse in the s+T group were higher than those of the s+T-NC group. (D) The cell proliferation ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group on the 2nd and 3rd day after transfection. (E) The cell migration ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group (magnification×200, scale bar: 100 μm). (F) The cell migration ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group (magnification×100). **P<0.01, ***P <0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Migration, Transfection, Over Expression, Plasmid Preparation, Produced

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: miR-335-3p acts as a competing endogenous RNA participating in the regulation of TEAD1 and AP000695.2 (A) The binding site of TEAD1 and miR-335-3p and the binding site of miR-335-3p and AP000695.2. (B) qPCR experiment showed that the expression of miR-335-3p in A549 cells was decreased after upregulating AP000695.2 expression. The expression of miR-335-3p in H1299 cells was increased after AP000695.2 was downregulated. (C) The expression of miR-335-3p in A549 and H299 cells was increased after the transfection of miR-335-3p mimics. (D) Western blot analysis showed that the protein expression of TEAD1 was decreased after the transfection with miR-335-3p mimics, and increased after the transfection with miR-335-3p inhibitor, compared with the negative control group. (E) Dual-luciferase reporter assay indicated that miR-335-3p has binding sites with TEAD1 and AP000695.2. **P<0.01, ***P<0.001. NS, no significance.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Binding Assay, Expressing, Transfection, Western Blot, Negative Control, Luciferase, Reporter Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: Silencing of AP000695.2 inhibits tumor growth and aerobic glycolysis in vivo (A) The qPCR experiment showed that the expression of si-AP000695.2 group is significantly lower than that of negative control group. (B) The tumor volumes at different time points showed that from the 6th day after injection, compared with the si-NC group, the si-AP000695.2 significantly inhibited the growth of transplanted tumor. (C) The uptake of 18F-FDG by tumor was decreased after the expression of AP000695.2 was decreased. The picture on the left showed the micro-PET images. The right picture showed the quantitative analysis results of SUVmax. (D) Immunohistochemistry (IHC) staining showed that the expression levels of GLUT1, HK2, PKM2, LDHA and TEAD1 in si-AP000695.2 group were lower than those in si-NC group. (magnification × 400, scale bar: 100 μm). **P<0.01, ***P<0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: In Vivo, Expressing, Negative Control, Injection, Micro-PET, Immunohistochemistry

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: Schematic diagram of the mechanisms of AP000695.2/miR-335-3p/TEAD1/GLUT1 axis in the glycolysis of LUAD AP000695.2 is involved in the mechanism of LUAD through functioning as a ceRNA to competitively sponge miR-335-3p, thereby regulating the expression of TEAD1. Then TEAD1 promotes glycolysis of lung adenocarcinoma by promoting GLUT1 transcription.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Expressing

Journal: RSC Chemical Biology

Article Title: A covalent inhibitor of the YAP–TEAD transcriptional complex identified by high-throughput screening †

doi: 10.1039/d3cb00044c

Figure Lengend Snippet:

Article Snippet: pCMV6-FLAG-TEAD1 , TEAD1 , N-Terminal FLAG , Origene RC215492.

Techniques: Plasmid Preparation, Luciferase

Journal: Nature Communications

Article Title: Analysis of chromatin accessibility uncovers TEAD1 as a regulator of migration in human glioblastoma

doi: 10.1038/s41467-018-06258-2

Figure Lengend Snippet: TEAD1 regulates GBM migration by modulating AQP4 expression. a Venn diagram depicts the intersection of genes with TEAD motifs and genes consistently downregulated in TEAD1KO cell lines and migration-deficient spheroids (striped area). AQP4 is the only one of 32 genes in this intersection found to be a direct TEAD1 binding target in vivo (highlighted in red). ATAC-seq set contains 2612 peak-annotated genes from E+GSC vs. E−GBM+NSPC differential accessibility analysis. Top RNA-seq set (overall targets) contains all 1648 significantly up or downregulated genes from TEAD1KO vs. Sham differential expression analysis in all samples from four different patient-derived GBM lines and three migration experiments ( n = 7; p adj. < 0.05; all log2(fold change) values of HGNC-annotated genes included). Bottom RNA-seq set (migratory targets) contains 865 significantly up or downregulated genes from TEAD1KO vs. Sham G-13063 spheroids from three independent migration experiments ( n = 3; p adj. < 0.05; log2(fold change) >1 or <−1). See also Supplementary Fig. . b Spheroid migration assay showing significant reversal of cell dispersion deficit at 30 h in TEAD1 knockout cells after overexpression of CDH11 or AQP4 (G-13063 line; laminin + PDL substrate; n = 3 wells. TEAD1KO + CDH11OE: ** p = 0.004; TEAD1KO + AQP4OE: ** p = 0.0015. Bars represent mean ± SEM). On right, migration area marked by red dash line in representative spheroids is shown. Scale bar = 75 μM. c Quantification of AQP4 expression by RT-qPCR. AQP4 is significantly upregulated in TEAD1KO cells after TEAD1 overexpression ( n = 6, 4 technical and 2 biological replicates; * p = 0.02 TEAD1KO + TEAD1OE vs. TEAD1KO) and is robustly expressed after exogenous lentivirus overexpression. Bars represent mean ± SEM

Article Snippet: For overexpression studies, the (Myc-DDK-tagged)-TEAD1, -AQP4, and CDH11 human ORF lentivirus constructs were transfected and purified from Lenti-X 293T cells (Origene RC215492L1, RC204693L1, and RC203810L1, respectively).

Techniques: Migration, Expressing, Binding Assay, In Vivo, RNA Sequencing, Quantitative Proteomics, Derivative Assay, Dispersion, Knock-Out, Over Expression, Quantitative RT-PCR