Journal: In Vivo

Article Title: miR-124 Targets EGFR and Attenuates Growth and Invasion in Bladder Cancer Cells

doi: 10.21873/invivo.14121

Figure Lengend Snippet: miR-124-3p inhibits bladder cancer cell progression through EGFR inactivation. (A) Cell viability of TSGH8301 and T24 cells treated with increasing concentrations of erlotinib, assessed by MTT assay. (B) Colony formation assay showing a significant reduction in clonogenic growth of TSGH8301 and T24 cells following erlotinib treatment. (C) Predicted binding sites of miR-124-3p on the 3’’ UTR of EGFR mRNA, as identified using the miRDB database. (D) Relative expression of miR-124-3p in TSGH8301 and T24 cells after erlotinib treatment, measured by qPCR. (E) Effects of miR-124-3p mimic on cell proliferation in TSGH8301 and T24 cells, evaluated via colony formation assay. Data are presented as mean ± standard error (SE) from at least three independent experiments (n=5). *p<0.05, **p<0.01 compared to control using one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Subsequently, cells on slides were incubated overnight at 4 ̊C with an unconjugated rabbit anti-EGFR primary antibody (1:300 in 1% BSA; #5605, Cell Signaling Technology).

Techniques: MTT Assay, Colony Assay, Binding Assay, Expressing, Control

Journal: In Vivo

Article Title: miR-124 Targets EGFR and Attenuates Growth and Invasion in Bladder Cancer Cells

doi: 10.21873/invivo.14121

Figure Lengend Snippet: miR-124-3p directly targets and inhibits EGFR expression in bladder cancer cells. (A) Western blot analysis showing reduced phosphorylation of EGFR (Tyr1068) in TSGH8301 and T24 cells transfected with the miR-124-3p mimic. (B) Quantification of EGFR levels showing over 20% inhibition by miR-124-3p, and p-EGFR (Tyr1068) levels showing over 70% inhibition by miR-124-3p. (C-D) Immunofluorescence staining for EGFR expression following treatment with miR-124-3p or erlotinib. Both treatments significantly reduced EGFR signal intensity. (E) Schematic representation of the wild-type (WT) and mutant (Mut) EGFR 3’’ UTR sequences used in the luciferase reporter assay. (F) Luciferase activity in cells co-transfected with miR-124-3p and either EGFR WT or EGFR Mut constructs. miR-124-3p suppressed luciferase activity in the WT group but not in the mutant group. Data are presented as mean ± standard error (SE) from at least three independent experiments (n=3). *p<0.05, **p<0.01 versus control using Student’s t test. Scale bar=100 μm.

Article Snippet: Subsequently, cells on slides were incubated overnight at 4 ̊C with an unconjugated rabbit anti-EGFR primary antibody (1:300 in 1% BSA; #5605, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Phospho-proteomics, Transfection, Inhibition, Immunofluorescence, Staining, Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Construct, Control

Journal: In Vivo

Article Title: miR-124 Targets EGFR and Attenuates Growth and Invasion in Bladder Cancer Cells

doi: 10.21873/invivo.14121

Figure Lengend Snippet: miR-124-3p suppresses invasion of bladder cancer cells through EGFR-mediated pathways. (A-B) Transwell invasion assay with Matrigel-coated chambers indicating suppressed invasive ability in both cell lines following miR-124-3p overexpression. (C) Western blot analysis and quantification of MMP2 (D), MMP9 (E), and VEGF-A (F) expression in TSGH8301 and T24 cells transfected with miR-124-3p. All three proteins, known to promote invasion and metastasis, were significantly downregulated by miR-124-3p. Data are presented as mean ± standard error (SE) (n=3). *p<0.05, **p<0.01 versus control using Student’s t test.

Article Snippet: Subsequently, cells on slides were incubated overnight at 4 ̊C with an unconjugated rabbit anti-EGFR primary antibody (1:300 in 1% BSA; #5605, Cell Signaling Technology).

Techniques: Transwell Invasion Assay, Over Expression, Western Blot, Expressing, Transfection, Control