Quercetin inhibits proliferation and migration and promotes apoptosis of gastric cancer cells by promoting phosphorylation-mediated YAP inactivation

doi:10.12122/j.issn.1673-4254.2026.02.15

Abstract

Abstract:

Objective To explore the molecular mechanisms by which quercetin regulates proliferation, migration, and apoptosis of gastric cancer cells. Methods Bioinformatics analysis was conducted to examine the differential expression of Hippo pathway protein yes-associated protein (YAP) between gastric cancer and normal tissues and its correlation with patient survival rates. CCK-8 assay was used to evaluate the effect of quercetin on viability of gastric cancer HGC-27 cells. The expression levels of YAP and phosphorylated YAP (p-YAP) in gastric cancer and normal gastric tissues were detected using Western blotting. In HGC-27 cells with YAP knockdown, the effects of quercetin treatment on cell proliferation, migration and apoptosis were assessed using EdU assay, colony formation assay, Transwell assay, and flow cytometry; the localization of YAP was determined using immunofluorescence staining. Rescue experiments were performed by overexpressing YAP in HGC-27 cells. Results YAP expression was markedly lower in gastric cancer tissues than in normal tissues, and its expression level was negatively correlated with patient survival rates. Quercetin significantly inhibited the survival of HGC-27 cells, promoted MST1 and LATS1 expression, and activated the Hippo signaling pathway, leading to phosphorylation-mediated inactivation of YAP, thereby reducing its nuclear translocation. Both YAP knockdown and quercetin treatment significantly suppressed proliferation and migration and promoted apoptosis of HGC-27 cells. Overexpression of YAP substantially attenuated the inhibitory effects of quercetin in HGC-27 cells. Conclusion Quercetin inhibits proliferation and migration and promotes apoptosis of gastric cancer cells by promoting phosphorylation-mediated inactivation of YAP.

Key words: Quercetin, Hippo signaling pathway, yes-associated protein, phosphorylation, gastric cancer

Yuhuang WANG, Wenrui WANG, Shujie CHENG, Yunlong FENG, Zhuo ZHANG, Qiying SHI, Yujia LI, Qianwen HU, Mingcai WU. Quercetin inhibits proliferation and migration and promotes apoptosis of gastric cancer cells by promoting phosphorylation-mediated YAP inactivation[J]. Journal of Southern Medical University, 2026, 46(2): 374-384.

Figures/Tables 9

Tab.1 Primer sequences for RT-qPCR

Primer	Forward (5'-3')	Reverse (5'-3')
LATS1	GTCGACGAGAGCAGATGGC	CACTTTCTCCTAGTGGCGGG
YAP	TGACCCTCGTTTTGCCATGA	GTTGCTGCGGTTGGAGTTG
CYR61	CAGGACTGTGAAGATGCGT	AGCCTGTAGAAGGGAAACGC
CTGF	GTCGAGTATGTACCGACGCG	GCAGGCACAGGTCTTGATGA
GAPDH	CGAGCCACATCGCTCAGACA	GTGGTGAAGACGCCAGTGGA

Fig.1 Expression levels of YAP and p-YAP in gastric cancer (GC) tissues and the correlation of YAP expression level with patient survival. A： Expression of YAP in GC (T) tissues and normal tissues (N) analyzed using the TCGA database. B： Correlation of YAP expression level with patient survival. C, D： Expression of YAP and p-YAP protein in normal gastric tissues and GC tissues. *P<0.05, ***P<0.001 vs Normal tissues.

Fig.2 Effects of YAP knockdown on cell proliferation, migration and apoptosis. A, B: YAP expression levels in HGC-27 cells with YAP knockdown. C: mRNA expression levels of YAP, CYR61 and CTGF in HGC-27 cells with YAP knockdown. D, E: EdU assays of HGC-27 cells with YAP knockdown （Scale bar=200 μm）. F, G: Colony formation assays of HGC-27 cells with YAP knockdown. H, I: Transwell migration assays of HGC-27 cells with YAP knockdown （Scale bar=100 μm）. J, K: Flow cytometric analysis of apoptosis of HGC-27 cells with YAP knockdown. *P<0.05, **P<0.01, ***P<0.001 vs si-NC group.

Fig.3 Quercetin inhibits viability of HGC-27 cells. **P<0.01, ***P<0.001 vs 0 μmol/L group.

Fig.4 Effects of quercetin on proliferation of HGC-27 cells. A, B:EdU assays for assessing proliferation of quercetin-treated HGC-27 cells (Scale bar=200 μm). C, D: Colony formation assays assessing proliferation of quercetin-treated HGC-27 cells. *P<0.05, **P<0.01 vs 0 μmol/L group.

Fig.5 Effects of quercetin on migration and apoptosis of HGC-27 cells. A, B: Transwell migration assays for assessing migratory ability of quercetin-treated HGC-27 cells (Scale bar=100 μm). C, D: Flow cytometry analysis of apoptosis of quercetin on quercetin-treated HGC-27 cells. *P<0.05, **P<0.01 vs 0 μmol/L group.

Fig.6 Effect of quercetin on Hippo signaling pathway protein expression in HGC-27 cells. A, B: Effect of 0-50 μmol/L quercetin on MST1, LATS1 and YAP protein expressions in HGC-27 cells. C, D： Effects of 50 μmol/L quercetin treatment for 24 h on MST1, p-MST1 LATS1, p-LATS1, YAP and p-YAP expression. E: Effect of 50 μmol/L quercetin on mRNA expression of LATS1, YAP and its downstream target genes CYR61 and CTGF in HGC-27 cells. F: Immunofluorescence staining for analyzing YAP nuclear and cytoplasmic expression in 50 μmol/L quercetin-treated HGC-27 cells (Scale bar=20 μm).*P<0.05, **P<0.01 vs 0 μmol/L group. #P<0.05, ##P<0.01 vs 0 h group.

Fig.7 Effect of YAP overexpression on YAP downstream genes in quercetin-treated gastric cancer cells. A, B: Western blotting for analyzing the inhibitory effects of quercetin on YAP in YAP-overexpressing HGC-27 cells. C: mRNA expression of YAP and its downstream target genes CYR61 and CTGF in HGC-27 cells in Control, Control+Que (50 μmol/L), oe-YAP and oe-YAP+Que (50 μmol/L) groups. **P<0.01, ***P<0.001 vs Control group.

Fig.8 Effect of YAP overexpression on proliferation, migration, and apoptosis of quercetin-treated gastric cancer cells. A, B: EdU assays for assessing proliferation of quercetin-treated HGC-27 cells overexpressing YAP. C, D: Colony formation assays for assessing proliferation of quercetin-treated HGC-27 cells overexpressing YAP. E, F: Transwell migration assays of quercetin-treated HGC-27 cells overexpressing YAP (Scale bar=100 μm). G, H: Flow cytometry for assessing apoptosis of quercetin-treated HGC-27 cells overexpressing YAP. *P<0.05, **P<0.01, ***P<0.001 vs Control group.

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