Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 1. NOEY2 inhibited tumour growth by suppressing angiogenesis in vivo. (A) Growth curves of human ovarian xenografts. NOEY2 stably transfected human SKOV-3 ovarian tumour cells, or empty vector (pcDNA3.1) stably transfected control cells were injected subcutaneously into nude mice to form tumours measuring 70 to 100 mm3. Tumour size was calculated every 3 days. *p<0.05; **p<0.01 versus control. (B) Tumours excised from control and NOEY2-overexpressed mice on day 28 of tumours measuring were photographed (left panel). Tumour weights were measured as the mean of three mice (n=5 per group) (right panel) ± SD. *p<0.05 versus control. (C) Sections of NOEY2-overexpressed and control tumours were stained with H&E (left panel), cell proliferation markers Ki67 (middle panel), and cleaved caspase-3 (right panel) to observe cell morphology and apoptotic cell death (indicated with the arrows), respectively. Bars=50 µm. (D, E) Whole-protein extracts were isolated from tumours of xenografted mice and analysed by western blotting using indicated antibodies. β-actin was used to verify equal loading amounts among samples. (F) Endothelial cells in paraffin-embedded tumour sections were stained using anti-CD31 antibodies (left panel). Bar=50 µm. *p<0.05 versus control. (G) Whole-protein extracts from tumours were evaluated the protein expression of VEGF, HIF-1α, phosphorylation of VEGFR-2 (Tyr-1175) and phosphorylation of VEGFR-2 (Tyr-951) by western blotting. β-actin and VEGFR-2 were used as the loading control. (H) Total cell lysates were prepared and detected by western blotting using anti-phospho-PI3K, anti-phospho-PDK1, anti-phospho-Akt, anti-phospho-mTOR, anti-phospho-TSC-2, anti-phospho-p70S6K and anti-phospho-GSK-3β antibodies. Total non-phosphorylated proteins (PI3K, PDK1, Akt, mTOR, TSC-2, p70S6K, and GSK-3β) were used to verify equal loading amounts among samples.
Biomolecules & Therapeutics 2021;29:506~518 https://doi.org/10.4062/biomolther.2021.121
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