Novel Anti-Angiogenic and Anti-Tumour Activities of the N-Terminal Domain of NOEY2 via Binding to VEGFR-2 in Ovarian Cancer

Abstract

The imprinted tumour suppressor NOEY2 is downregulated in various cancer types, including ovarian cancers. Recent data suggest that NOEY2 plays an essential role in regulating the cell cycle, angiogenesis and autophagy in tumorigenesis. However, its detailed molecular function and mechanisms in ovarian tumours remain unclear. In this report, we initially demonstrated the inhibitory effect of NOEY2 on tumour growth by utilising a xenograft tumour model. NOEY2 attenuated the cell growth approximately fourfold and significantly reduced tumour vascularity. NOEY2 inhibited the phosphorylation of the signalling components downstream of phosphatidylinositol-3’-kinase (PI3K), including phosphoinositide-dependent protein kinase 1 (PDK-1), tuberous sclerosis complex 2 (TSC-2) and p70 ribosomal protein S6 kinase (p70S6K), during ovarian tumour progression via direct binding to vascular endothelial growth factor receptor-2 (VEGFR-2). Particularly, the N-terminal domain of NOEY2 (NOEY2-N) had a potent anti-angiogenic activity and dramatically downregulated VEGF and hypoxia-inducible factor-1α (HIF-1α), key regulators of angiogenesis. Since no X-ray or nuclear magnetic resonance structures is available for NOEY2, we constructed the three-dimensional structure of this protein via molecular modelling methods, such as homology modelling and molecular dynamic simulations. Thereby, Lys15 and Arg16 appeared as key residues in the N-terminal domain. We also found that NOEY2-N acts as a potent inhibitor of tumorigenesis and angiogenesis. These findings provide convincing evidence that NOEY2-N regulates endothelial cell function and angiogenesis by interrupting the VEGFR-2/PDK-1/GSK-3β signal transduction and thus strongly suggest that NOEY2-N might serve as a novel anti-tumour and anti-angiogenic agent against many diseases, including ovarian cancer.

Snail Promotes Cancer Cell Proliferation via Its Interaction with the BIRC3

“…Oligonucleotides including an RNA interference (RNAi) sequence [Snail: 5’- GCC UUC AAC UGC AAA UAC U-3’ (sense) and 5’-AGU AUU UGC AGU UGA AGG C-3’ (antisense); BIRC3, 5’-UAA GGG AAG AGG AGA GAG AA-3’ (sense) and 5’-UUC UCU CUC CUC UUC CCU UA-3’ (antisense)] were synthesized using an RNAi construction kit (Ambion, Austin, TX, USA), and were then transfected with Lipofectamine ® 2000 reagent (Invitrogen) according to the manufacturer’s protocols ( Rho et al ., 2021b ). The following unrelated non-specific scrambled oligonucleotide was used as a control: [Snail, 5’-TAT TGC CTA GCA TTA CGT T-3’ (sense) and 5’-GTA TTG CCT AGC ATT ACG T-3’ (antisense); BIRC3, 5’-CCT GGT AGC AGC GAG TGA G-3’ (sense) and 5’-CTC ACT CGC TGC TAC CAG G-3’ (antisense)].…”

PRR16/Largen Induces Epithelial-Mesenchymal Transition through the Interaction with ABI2 Leading to the Activation of ABL1 Kinase

“…Total RNA was prepared using TRIzol ® RNA Isolation Reagents (Invitrogen) according to the manufacturer’s instructions ( Rho et al ., 2021b ). Reverse transcription was performed with a First Strand cDNA Synthesis kit (Promega, Madison, WI, USA).…”

HPV-18 E7 Interacts with Elk-1 Leading to Elevation of the Transcriptional Activity of Elk-1 in Cervical Cancer

“…Yeast two-hybrid screening was performed using the Matchmaker LexA two-hybrid system (Clontech, Mountain View, CA, USA) according to the manufacturer’s instructions ( Rho et al ., 2021b ). Briefly, E7 cDNA was cloned into the pGilda bait vector as described above and human ovary cDNA library used as prey vector.…”