Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: The COL1A2 promoter is significantly regulated by an enhancer-like signature. (a) COL1A2 promoter modification level searched by UCSC browser. (b, c) ChIP-seq data of H3K27ac in GC and normal gastric tissues were downloaded from the GEO database and analyzed for enrichment in the COL1A2 promoter. (d) ChIP-qPCR analysis of COL1A2 promoter histone H3K27ac modification levels in AGS and MKN-45 cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. IgG).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Modification, ChIP-sequencing

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: EP300 promotes COL1A2 expression through an H3K27ac-dependent manner. GC cells were treated with HAT inhibitor or DMSO. (a) ChIP-qPCR analysis of the H3K27ac modification level of COL1A2 promoter. (b) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (c) The protein expression of COL1A2 in parental cells by western blot. (d) UCSC browser analysis of the binding of EP300 to the promoter of COL1A2. GC cells were transfected with oe-NC or oe-EP300. (e) ChIP-qPCR analysis of the H3K27ac modification level of COL1A2 promoter. (f) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (g) The protein expression of COL1A2 in parental cells by western blot. (h) Colocalization of TWIST1 with EP300 in AGS and MKN-45 cells by double-labeled immunofluorescence staining. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. DMSO or oe-NC).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Expressing, Modification, Quantitative RT-PCR, Western Blot, Binding Assay, Transfection, Labeling, Immunofluorescence, Staining

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Overexpression of COL1A2 promotes resistance of parental cells to apatinib. GC parental cells were transfected with oe-NC or oe-COL1A2. (a) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (b) The protein expression of COL1A2 in parental cells by western blot. (c) The IC50 values of parental cells by CCK-8 assay. (d) The number of colonies formed by parental MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in parental MKN-45 and AGS cells. (f) TUNEL analysis of the number of apoptotic bodies in parental MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. oe-NC).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, TUNEL Assay

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Knockdown of COL1A2 inhibits the sensitivity of drug-resistant cells to apatinib. GC-resistant cells were transfected with Scr or sh-COL1A2 #1 or #2. (a) The mRNA expression of COL1A2 in resistant cells by RT-qPCR. (b) The protein expression of COL1A2 in resistant cells by western blot. (c) The IC50 values of resistant cells by CCK-8 assay. (d) The number of colonies formed by resistant MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in resistant MKN-45 and AGS cells. (f) TUNEL analysis of the number of apoptotic bodies in resistant MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. Scr).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, TUNEL Assay

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Knockdown of TWIST1 or EP300 reduces resistance to apatinib in parental cells overexpressing COL1A2. GC parental cells transfected with oe-COL1A2 were further transfected with sh-TWIST1 or sh-EP300. (a) The mRNA expression of TWIST1, EP300, and COL1A2 in parental cells by RT-qPCR. (b) The protein expression of TWIST1, EP300, and COL1A2 in parental cells by western blot. (c) The IC50 values of parental cells by CCK-8 assay. (d) The number of colonies formed by parental MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in parental MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. oe − COL1A2 + Scr).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry