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Article Open Access

RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability

  • Authors:
    • Siyang Peng
    • Yidong Chen
    • Jieke Wu
    • Xiaodong Huang
    • Linjie Hong
    • Yanci Xie
    • Yuting Lei
    • Xiangyang Wei
    • Ping Yang
    • Jieming Zhang
    • Qiong Yang
    • Guangnan Liu
    • Aimin Li
    • Side Liu
    • Jiaying Li
    • Weiyu Dai
    • Yanfeng Hu
    • Jing Wang
    • Jing Xiong
    • Jide Wang
  • View Affiliations / Copyright

    Affiliations: Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Gastroenterology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian 362000, P.R. China, Department of Gastroenterology, The Key Laboratory of Advanced Interdisciplinary Studies Center, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, P.R. China, Department of Gastroenterology and Hepatology, Guangdong Provincial People's Hospital Guangdong Academy of Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510080, P.R. China, Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Pathology, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510163, P.R. China
    Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 50
    |
    Published online on: February 26, 2026
       https://doi.org/10.3892/ijo.2026.5863
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Abstract

DAZ‑associated protein 1 (DAZAP1), an RNA‑binding protein and modulator of alternative splicing, participates in tumorigenesis. However, the potential oncogenic function and mechanism of DAZAP1 in gastric cancer (GC) are unknown. Gene expression analysis, including mRNA and protein level assessment by reverse transcription‑quantitative PCR and western blotting, respectively, immunofluorescence, immunohistochemistry, in situ hybridization assays, tissue microarray, RNA immunoprecipitation and sequencing and mRNA stability assay were performed, as well as colony formation, EdU, wound healing, migration and invasion assays of GC cells. DAZAP1 displayed a significant upregulation in GC cells and served as an oncogene, as demonstrated by its overexpression promoting colony formation, EdU incorporation, wound healing, migration and invasion, and its knockdown suppressing these malignant phenotypes. Additionally, DAZAP1 upregulation was positively correlated with tumor progression and poor survival in individuals with GC. Functionally, DAZAP overexpression promoted proliferation, epithelial‑mesenchymal transition (EMT) and migration/invasion of GC cells. Mechanistically, DAZAP1 physically bound NOTCH1 or JAG1 mRNA to regulate its stability. In addition, overexpression of DAZAP1 facilitated NOTCH1‑ and/or JAG1‑mediated migration via EMT in GC cells. Changes in NOTCH1 or JAG1 expression were positively correlated with DAZAP1 expression when DAZAP1 was silenced or enhanced in GC. Finally, DAZAP1 modulated the activation of the NOTCH/JAG1 signaling pathway. DAZAP1 expression facilitated migration/invasion and mediated the stabilization of NOTCH1 or JAG1 mRNA, suggesting they may participate in GC progression.
View Figures

Figure 1

DAZAP1 is upregulated in GC. (A)
Differentially expressed genes in the three Gene Expression Omnibus
datasets and the RBPmap website. (B) Relative expression of DAZAP1
in 10 GC and their adjacent N tissues was examined by western blot
assay. β-tubulin was used as the internal control. (C) Expression
of DAZAP1 protein in six human GC and normal mucous cell line
GES-1. (D) Representative staining of DAZAP1 intensity using IHC.
(Ea) Representative gastric tissue from two cancerous and
non-cancerous patients. Expression of DAZAP1 in normal and
malignant human gastric tissue was detected by tissue microarray.
The arrows indicate the cells with epithelial-to-mesenchymal
transition) morphological alterations. (Eb) Expression of DAZAP1
was determined in N and T gastric tissue. Scale bar, 100 μm.
Kaplan-Meier survival analysis of overall survival in (Fa) all
patients and those with (Fb) early and (Fc) late stage GC
(according to DAZAP1 expression. The log-rank test was used to
calculate P-values. ****P<0.001. DAZAP, deleted in
azoospermia-associated protein 1; GC, gastric cancer; N, normal
tissue; T, tumor tissue; PPARGC1A, peroxisome
proliferator-activated receptor gamma coactivator 1 alpha; CIRBP,
cold inducible RNA-binding protein; PABPC4, poly(A)-binding protein
cytoplasmic; FC, fold-change; IHC, immunohistochemistry.

Figure 2

Functional analysis of DAZAP1 in GC
cells in vitro. (A) Colony formation assay was performed to
detect proliferative capability at 48 h. (B) EdU assay was used to
determine the viability of GC cells. Red, EdU-positive cells; blue,
Hoechst33258 (total cells). Scale bar, 100 μm. (C) Motility
was determined wound healing assay. (D) Migration and (E) invasion
assay were performed following transfection in GC cells. Scale bar,
150 μm. ***P<0.01, ****P<0.001
vs. vector; ###P<0.01, ####P<0.001
Scr-siRNA vs. DAZAP1-siRNAp. DAZAP1, deleted in
azoospermia-associated protein 1; GC, gastric cancer; scr,
scramble; si, small interfering; p, pool.

Figure 3

DAZAP1 induces EMT in GC cells. (A)
Phase-contrast microscopy in MKN-45 cells. Scale bar, 50 μm.
(B) F-actin expression and localization were analyzed by
immunocytochemistry. The cytoskeleton and nuclei were stained with
rhodamine-phallotoxin (red) and Hoechst33258 (blue). (C)
Immunofluorescence staining for E-cadherin (green) and vimentin
(red) in MKN-45 cells. Scale bar, 20 μm. (D) EMT biomarkers,
including E-cadherin, N-cadherin and vimentin, were detected by
western blotting in GC cells. (E) Representative metastatic tumors
in the lungs from mice (n=4/group). Scale bar, 1 cm. (F) Number of
metastatic tumors in the lung was counted. (G) Hematoxylin and
eosin staining of lung sections of mice. (H) Immunohistochemical
staining of vimentin. Scale bar, 100 μm.
****P<0.001. DAZAP, deleted in azoospermia-associated
protein 1; EMT, epithelial-mesenchymal transition; GC, gastric
cancer; scr, scramble; si, small interfering; sh, short
hairpin.

Figure 4

DAZAP1 binds NOTCH1 or JAG1 mRNA in
GC cells. (A) DAZAP1-binding motif was predicted using the RBPmap
database (rbpmap.technion.ac.il/). (B) Overlap between
DAZAP1-interacting genes identified using RIP-seq and EMT-related
genes from the EMT database (emtome.org).
(C) Quantification of target genes involved in the pathways in the
cancer signaling pathway, including NOTCH1 and JAG1, according to
KEGG analysis in the WebGestalt database (webgestalt.org/). (D) RT-qPCR analysis of the mRNA
levels of NOTCH1 and JAG1 following DAZAP1 knockdown in AGS and
MKN-45 cells. RT-qPCR analysis of (E) NOTCH1 and (F) JAG1 with
anti-DAZAP1 and IgG control antibodies in AGS cells. (G) NOTCH1 and
(H) JAG1 mRNA t1/2 in DAZAP1 knockdown AGS cells. n=3.
(I) Verification of full-length or truncated DAZAP1 plasmids using
western blotting and anti-FLAG antibody. RIP assay analysis of (J)
NOTCH1 and (K) JAG1 mRNA binding to the DAZAP1 structure domains.
***P<0.01 and ****P<0.001 vs.
scr-siRNA. RRM, RNA recognition motif; CTD, C-terminal domain;
DAZAP1, deleted in azoospermia-associated protein 1; RIP-seq, RNA
Immunoprecipitation-sequencing; EMT, Epithelial-Mesenchymal
Transition; KEGG, Kyoto Encyclopedia of Genes and Genomes; RT-q,
Reverse Transcription quantitative real-time; t1/2,
half-life; scr, scramble; si, small interfering; F, Fragment; FL,
Full-Length; aa, amino acid.

Figure 5

DAZAP1 regulates NOTCH1 or JAG1
expression to affect migration/invasion via epithelial-mesenchymal
transition in GC cells. The wound healing assay in MKN-45 cells
confirmed that DAZAP1 regulated (A) NOTCH1(A) and JAG1. (B) Scale
bar, 100 μm. The Transwell migration assay confirmed that
DAZAP1 regulates NOTCH1(C) and JAG1 (D) in MKN-45 and AGS cells,
Scale bar, 150 μm. Expression of epithelial marker
E-cadherin and mesenchymal markers vimentin and N-cadherin was
detected by western blotting with β-tubulin as the internal control
in AGS cells. The western blotting in AGS cells confirmed that
DAZAP1 regulates NOTCH1 (E) and JAG1 (F). ***P<0.01,
****P<0.001. DAZAP1, deleted in
azoospermia-associated protein 1; GC, gastric cancer; scr,
scramble; si, small interfering.

Figure 6

DAZAP1 promotes NOTCH1 and JAG1
expression to alter migration/invasion via epithelial-mesenchymal
transition in gastric cancer cells. (A) Cell migration was tested
using a monolayer wound healing assay. Scale bar, 100 μm.
(B) Migration and (C) invasion assay. Scale bar, 150 μm. (D)
Epithelial (E-cadherin) and mesenchymal markers (N-cadherin,
vimentin) were detected by western blotting in AGS and MKN-45
cells. (E) Lung samples were collected from mice injected with AGS
cells via the tail vein. Scale bar, 1 cm (F) Lung metastatic
nodules were counted. (G) Hematoxylin and eosin and (H) vimentin
staining were conducted on lung samples from these mice. Scale bar,
100 μm. ****P<0.001. DAZAP1, deleted in
azoospermia-associated protein 1; scr, scramble; si, small
interfering; sh, short hairpin.

Figure 7

DAZAP1 expression is positively
correlated with NOTCH1 or JAG1 mRNA expression in GC tissues. (A)
Expression of DAZAP1 and (B) Expression levels of NOTCH1 in 10
paired N and GC tissue specimens were detected by qPCR.
***P<0.01, ****P<0.001 (C) Expression
levels of JAG1 in 10 paired N and GC tissue specimens were detected
by qPCR. ****P<0.001 (D) Correlation between DAZAP1
and (D) NOTCH1 in 10 GC tissue samples. (E) Correlation between
DAZAP1 and JAG1 in 10 GC tissue samples. (F) Gene expression in N
and GC tissue was evaluated using immunohistochemistry and in
situ hybridization. (G) Abnormal expression of DAZAP1 modulated
the NOTCH1 signaling pathway, as detected by western blot analysis.
(H) Signal transduction mechanism of DAZAP1. Scale bar, 100
μm. DAZAP1, deleted in azoospermia-associated protein 1; GC,
gastric cancer; N, normal; T, tumor; HES, Hairy and Enhancer of
Split 1; NICD, Notch intracellular domain; C, case; scr, scramble;
si, small interfering.
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Copy and paste a formatted citation
Spandidos Publications style
Peng S, Chen Y, Wu J, Huang X, Hong L, Xie Y, Lei Y, Wei X, Yang P, Zhang J, Zhang J, et al: RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability. Int J Oncol 68: 50, 2026.
APA
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y. ... Wang, J. (2026). RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability. International Journal of Oncology, 68, 50. https://doi.org/10.3892/ijo.2026.5863
MLA
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y., Lei, Y., Wei, X., Yang, P., Zhang, J., Yang, Q., Liu, G., Li, A., Liu, S., Li, J., Dai, W., Hu, Y., Wang, J., Xiong, J., Wang, J."RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability". International Journal of Oncology 68.5 (2026): 50.
Chicago
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y., Lei, Y., Wei, X., Yang, P., Zhang, J., Yang, Q., Liu, G., Li, A., Liu, S., Li, J., Dai, W., Hu, Y., Wang, J., Xiong, J., Wang, J."RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability". International Journal of Oncology 68, no. 5 (2026): 50. https://doi.org/10.3892/ijo.2026.5863
Copy and paste a formatted citation
x
Spandidos Publications style
Peng S, Chen Y, Wu J, Huang X, Hong L, Xie Y, Lei Y, Wei X, Yang P, Zhang J, Zhang J, et al: RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability. Int J Oncol 68: 50, 2026.
APA
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y. ... Wang, J. (2026). RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability. International Journal of Oncology, 68, 50. https://doi.org/10.3892/ijo.2026.5863
MLA
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y., Lei, Y., Wei, X., Yang, P., Zhang, J., Yang, Q., Liu, G., Li, A., Liu, S., Li, J., Dai, W., Hu, Y., Wang, J., Xiong, J., Wang, J."RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability". International Journal of Oncology 68.5 (2026): 50.
Chicago
Peng, S., Chen, Y., Wu, J., Huang, X., Hong, L., Xie, Y., Lei, Y., Wei, X., Yang, P., Zhang, J., Yang, Q., Liu, G., Li, A., Liu, S., Li, J., Dai, W., Hu, Y., Wang, J., Xiong, J., Wang, J."RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability". International Journal of Oncology 68, no. 5 (2026): 50. https://doi.org/10.3892/ijo.2026.5863
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