Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Modulation of RASD2 by miRNA ‐485‐5p Drives Thyroid Cancer Progression and Metastasis

doi: 10.1002/kjm2.70028

Figure Lengend Snippet: miR‐485‐5p regulation of RASD2 in thyroid cancer. (A) Venn diagram showing overlapping miRNA predictions from three databases (TargetScan, miRDB, StarBase). (B) Sequence alignment between miR‐485‐5p and RASD2 3′‐UTR showing predicted binding site. (C) Dual‐luciferase reporter assay results in IHH4 and TPC‐1 cells transfected with wild‐type or mutant RASD2 3′‐UTR constructs and miR‐485‐5p mimic or negative control. (D) Western blot analysis of RASD2 protein levels in cells transfected with miR‐485‐5p mimic or negative control. (E) TCGA database analysis of miR‐485‐5p expression in thyroid cancer vs. normal tissues. (F) miR‐485‐5p expression in paired tumor‐normal tissues by qRT‐PCR ( n = 96 pairs). (G) Correlation analysis between miR‐485‐5p and RASD2 expression levels in thyroid cancer tissues. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After 48 h, luciferase activities were measured using the Dual‐Luciferase Reporter Assay System (HY‐K1013, MedChemExpress).

Techniques: Sequencing, Binding Assay, Luciferase, Reporter Assay, Transfection, Mutagenesis, Construct, Negative Control, Western Blot, Expressing, Quantitative RT-PCR

Journal: Journal of Gastrointestinal Oncology

Article Title: Nestin overexpression reduces the sensitivity of gastric cancer cells to trastuzumab

doi: 10.21037/jgo-22-1048

Figure Lengend Snippet: Influence of Nestin on the activation of Nrf2/HO-1 and HER2/AKT/ERK1/2 of HER2-positive GC cells. The two cell lines received transfection by lentiviral-Nestin or lentiviral-empty, and then added to 10 μg/mL for 48 h. (A,B) Nestin mRNA level in MKN45 and NCI-N87 cells was detected by qRT-PCR. (C,D) WB was adopted to determine the protein level of Nestin, Nrf2, HO-1, NQO1, GST, AKT, ERK1/2, and HER2, in the MKN45 and NCI-N87 cells. (E,F) The cells were exposed to ARE luciferase reporter. 36 h post transfection, the luciferase activity was determined. Data are presented as the means ± SD of three independent experiments. *, P<0.05; **, P<0.01; ***, P<0.001. NTC, non-treated control; TRA, trastuzumab; TRA + NC, mice inoculated with cell infected with lentiviral-negative control were treated with TRA; TRA + Nestin, mice inoculated with cell infected with lentiviral-Nestin were treated with TRA; HO-1, hemeoxygenase-1; NQO1, NAD(P)H: quinone oxidoreductase-1; GST, glutathione S-transferase; AKT, protein kinase B; ERK1/2, extracellular regulated kinase 1/2; HER2, human epidermal growth factor receptor 2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ARE, antioxidant response element; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WB, Western blotting; SD, standard deviation.

Article Snippet: After a day of culture, the cells were treated with 200 ng of ARE luciferase reporter plasmids using OriGene transfection reagent (Company, City, Country).

Techniques: Activation Assay, Transfection, Quantitative RT-PCR, Luciferase, Activity Assay, Control, Infection, Negative Control, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation