YAP promotes the proliferation of neuroblastoma cells through decreasing the nuclear location of p27^Kip1 mediated by Akt

Abstract

Objective We aimed to investigate the roles and underlying mechanisms of YAP in the proliferation of neuroblastoma cells. Methods The expression level of YAP was evaluated by Western blotting and immunocytochemistry. Cell viability, cell proliferation and growth were detected by CCK‐8, PH3 and Ki67 immunostaining, and the real‐time cell analyser system. The nuclear and cytoplasmic proteins of p27Kip1 were dissociated by the nuclear‐cytosol extraction kit and were detected by Western blotting and immunocytochemistry. mRNA levels of Akt, CDK5 and CRM1 were determined by qRT‐PCR. Results YAP was enriched in SH‐SY5Y cells (a human neuroblastoma cell line). Knock‐down of YAP in SH‐SY5Y cells or SK‐N‐SH cell line (another human neuroblastoma cell line) significantly decreased cell viability, inhibited cell proliferation and growth. Mechanistically, knock‐down of YAP increased the nuclear location of p27Kip1, whereas serum‐induced YAP activation decreased the nuclear location of p27Kip1 and was required for cell proliferation. Meanwhile, overexpression of YAP in these serum‐starved SH‐SY5Y cells decreased the nuclear location of p27Kip1, promoted cell proliferation and overexpression of p27Kip1 in YAP‐activated cells inhibited cell proliferation. Furthermore, knock‐down of YAP reduced Akt mRNA and protein levels. Overexpression of Akt in YAP‐downregulated cells decreased the nuclear location of p27Kip1 and accelerated the proliferation of SH‐SY5Y cells. Conclusions Our studies suggest that YAP promotes the proliferation of neuroblastoma cells through negatively controlling the nuclear location of p27Kip1 mediated by Akt.

Hydrogels for 3D Neural Tissue Models: Understanding Cell-Material Interactions at a Molecular Level

“…The gene analysis of p27 /Kip1 showed a significant twofold upregulation by Days 7 and 10, when compared to Day 3 ( Figure 5E). These observations agree with other reports linking increased p27 /Kip1 expression with reduced YAP expression (Shen et al, 2020).…”

Astrocytic YAP protects the optic nerve and retina in an experimental autoimmune encephalomyelitis model through TGF-β signaling

“…Since YAP is involved in proliferation of various cells, such as astrocytes, glioma cells and olfactory ensheathing cells 39 , 43 , 52 , to further explore whether the inflammatory infiltration in YAP GFAP -CKO mice was caused by the proliferation of astrocytes and microglia, double immunostaining of Ki67/GFAP, PH3/GFAP, Ki67/Iba1, and PH3/Iba1 (Ki67 and PH3, cell markers of proliferation) were performed in optic nerve of YAP f/f EAE and YAP GFAP -CKO EAE mice. As shown in Figure 4 A-D, the proliferation of astrocytes was decreased in optic nerve of YAP GFAP -CKO EAE mice, compared with that in YAP f/f EAE mice.…”

Luteolin Ameliorates Experimental Pulmonary Arterial Hypertension via Suppressing Hippo-YAP/PI3K/AKT Signaling Pathway

“…Among them, AKT is the most probable candidate (Kudryashova et al, 2016). YAP can directly activate the expression of Pik3cb through TEAD, promoting the transcription of PI3K and then activating AKT (Lin et al, 2015;Shen et al, 2020). PI3K/AKT signal plays an important role in regulation of cell migration, proliferation and survival (Yu & Cui, 2016).…”

“…The latest research also has proved that luteolin is a new type of YAP inhibitor (Cao et al, 2020). At the same time, YAP promotes cell proliferation by activating the AKT pathway (Lin et al, 2015;Shen et al, 2020). YAP and PI3K/ AKT signaling pathways play an important role in regulating the proliferation and migration of PASMCs (Teng et al, 2003;Kudryashova et al, 2016;Huang et al, 2017;Zhang et al, 2018).…”