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10.1007/s12672-024-01354-w
Research
hnRNPA1 promotes the metastasis and proliferation of gastric cancer cells through WISP2-guided Wnt/β-catenin signaling pathway
Jiang Chenyang 1
Xu Dengfei 1
Feng Hao 1
Ren Zirui 1
Li Xiang 1
Chen Yuming 1
Yu Jifeng yujifengzzu@163.com

2
Cang Shundong shundongcang@zzu.edu.cn

1
1 https://ror.org/03f72zw41 grid.414011.1 0000 0004 1808 090X Department of Oncology Henan Key Laboratory for Precision Medicine in Cancer, Henan Provincial People’s Hospital, No 7, Weiwu Rd, Zhengzhou, 450003 Henan China
2 https://ror.org/056swr059 grid.412633.1 Department of Hematology, The First Affiliated Hospital of Zhengzhou University, No. 1, Jianshe East Rd, Zhengzhou, 450003 Henan China
19 9 2024
19 9 2024
12 2024
15 46527 6 2024
16 9 2024
© The Author(s) 2024
2024
https://creativecommons.org/licenses/by-nc-nd/4.0/ Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
The main cause of gastric cancer (GC)-related death is due to malignant cell unregulated distant metastasis and proliferation. Heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) has been shown to play an important role in carcinogenesis and the development of metastasis in several tumors. However, its downstream regulatory mechanism in GC is not well defined. Our study aims to investigate the function and regulatory mechanism of hnRNPA1 in GC. We analyzed the differential expression of hnRNPA1 in gastric cancer and paired adjacent normal tissues in the TCGA database. Kaplan-Meier analysis was employed for survival assessment. The expressions of hnRNPA1 in GC cells were measured by qRT-PCR and Western blot. Transwell assay, CCK8 and colony formation assay were used to detect the effect of hnRNPA1 on the metastasis and proliferation ability of GC cells. Additionally, Western blotting was performed to examine the expression of proteins related to the Wnt/β-catenin signaling pathway as well as epithelial-mesenchymal transition (EMT), while further investigations were carried out to explore potential regulatory mechanisms. The results showed that hnRNPA1 was highly expressed differentially in GC over normal gastric tissue. Knocking down hnRNPA1 inhibited the metastasis and proliferation of human gastric cancer cells. Overexpression of hnRNPA1 significantly enhanced the metastatic potential and proliferative capacity of human GC cells. Further mechanism exploration revealed that knocking down hnRNPA1 inhibited the Wnt/β-catenin signaling pathway and WNT1 inducible signaling pathway protein-2 (WISP2), an activator of the Wnt/β-catenin signaling pathway. Whereas overexpression of hnRNPA1 had the opposite effects. Our results demonstrated that hnRNPA1 promoted metastasis and proliferation of GC cells by activating Wnt/β-catenin signaling pathway via WISP2. hnRNPA1 may serve as a potential biomarker and novel therapeutic targets for GC.

Supplementary Information

The online version contains supplementary material available at 10.1007/s12672-024-01354-w.

Keywords

AGS
HnRNPA1
WISP2
Wnt/β-catenin signaling pathway
Gastric cancer
Mechanism of cancer progression
shundongcang,Natural Science Foundation of Henan Province22230042050 science and technology project of Henan Province 212102310161 LHGJ20220051 issue-copyright-statement© Springer Science+Business Media, LLC 2024
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pmcIntroduction

Gastric cancer is the 5th leading cause of cancer-related mortality worldwide [following lung, colorectal, liver and breast cancers] accounting for 6.8% of all cancer deaths, with a particularly significant higher incidence rate in regions like China and other eastern Asian countries [1]. Due to the insidious and often nonspecific symptoms associated with this malignancy, unfortunately, greater than 60% of the patients were diagnosed with reginal or distant metastasis in the unscreened population [2]. Current treatment modalities for gastric cancer encompass a combination of perioperative chemotherapy, surgical intervention and/or radiation therapy, depending on the stage and extent of the disease. While there have been notable progresses in cancer research and treatment options, the five-year survival rate for patients with distant metastasis remains less than 10% [3]. This underscores the critical need for continued research to understand the molecular mechanism of gastric cancer cell metastasis or proliferation to improve outcomes and ultimately reduce the burden of this devastating disease.

Heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) is an RNA-binding protein renowned for its involvement in various facets of RNA regulation, encompassing mRNA packaging, alternative splicing and translational regulation [4]. Within the hnRNPs family, it stands as one of the most extensively studied proteins. High expression of hnRNPA1 is commonly found in many types of tumors, including lung cancer, liver cancer, colorectal adenocarcinoma, and pancreatic cancer [5–8], and is usually associated with metastasis or proliferation of tumor cells [9, 10]. Knockdown of hnRNPA1 effectively inhibited the metastasis or proliferation of pancreatic cancer, chronic lymphocytic leukemia, lung cancer, and gastric cancer [11–14]. However, up to now, the downstream mechanism of action of hnRNPA1 in GC remains to be further elucidated.

Wnt/β-catenin signaling pathway is one of the main signaling pathways that can be initiated by Wnt signaling [15]. Numerous studies have shown that Wnt/β-catenin signaling pathway is involved in a variety of pathophysiological processes, including cell proliferation, motility and angiogenesis [16, 17], and participates in the occurrence and development of gastric cancer [18, 19]. Evidence suggests a connection between WNT1 inducible signaling pathway protein-2 (WISP2) and β-catenin in gastric cancer [20]. WISP2 has also been shown to degrade the β-catenin degradation complex and promote the accumulation of β-catenin in the cytoplasm [21]. WISP2 stands as a significant member within the cysteine-rich glycosylated signaling protein (CCN) family involved in oncogenesis [22, 23]. It is generally believed that Wnt/β-catenin signaling pathway leads to the activation of the epithelial mesenchymal transition (EMT) cascade [24]. A substantial body of research underscores that the occurrence of EMT heralds the onset of tumor progression such as tumor migration and invasion [25–28].

In this study, we explored the expression of hnRNPA1 in GC cell lines and its biological effects on GC cell metastasis and proliferation. We further investigated and demonstrated that hnRNPA1 activates Wnt/β-catenin signaling pathway through WISP2 to promote GC cell metastasis and proliferation.

Materials and methods

Database analysis

To validate hnRNPA1 as a reproducible biomarker for GC, we downloaded the data of hnRNPA1 in gastric cancer and adjacent normal tissues from the TCGA database, integrated the data with Perl program algorithm, and finally obtained 33 paired data and analyzed the data with paired samples t-test technology. We conducted an analysis of the relationship between the quantity of hnRNPA1 expression and distant metastasis using the Biomarker Exploration of Solid Tumors (https://rookieutopia.com/app_direct/BEST/) platform on the GSE database. Survival analysis was employed to assess overall survival (OS) and post progression survival (PPS) using Kaplan Meier-Plotter (https://kmplot.com/analysis/).

Cell lines and cell culture

Human GC cell lines (AGS, HGC27 and MKN45) and human normal gastric mucosal epithelial cells (GES-1) were purchased from Nanjing Kebai Biotechnology Co. LTD. (Nanjing, China). These cells were seeded in RPMI-1640 medium (Invitrogen, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, CA, USA) and cultured at 37 °C in a cell incubator with 5% CO2.

Cell transfection

AGS cells were plated in 6-well culture plates, and transfection was facilitated using the Lipofectamine 3000 Transfection Kit (Invitrogen, CA, USA) for the introduction of Small interfering RNA (siRNA) and plasmid into the cells. The siRNA was acquired from Ribobio (Guangzhou, China). The pcDNA3.1 plasmid, pcDNA3.1-hnRNPA1 plasmid and pcDNA3.1-WISP2 plasmid were obtained from TSINGKE Biological Technology (Beijing, China). Cells were harvested 24 to 48 h after transfection for analysis. The sequences used above are shown in Supplementary Table S1.

RNA extraction and quantitative real-time PCR (qRT–PCR) assay

RNA extraction was performed utilizing the TRIZOL reagent (Invitrogen, CA, USA) according to the product specification. One microliter of RNA was subjected to reverse transcription into cDNA employing the Primescript RT reagent kit (Vazyme Biotech, Nanjing, China). The quantification of cDNA was carried out through qRT-PCR, and the data were recorded using FS Universal SYBR Green Master (Roche, Basel, Switzerland) and the Applied Biosystems 7500 instrument. β-actin was used as an internal reference. The primers are listed in Supplementary Table S2.

Western blot assay

The related proteins in this study were extracted on ice using Radio Immunoprecipitation Assay (RIPA) lysis buffer (Beyotime, Shanghai, China). Protein concentrations were determined using the BCA Protein Assay kit (Thermo Scientific, CA, USA). Equal quantities of proteins were loaded on 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) to electrophorese and subsequently transferred onto polyvinylidene difluoride (PVDF) membranes (Merck, Darmstadt, Germany). Membranes were trimmed appropriately and incubated with primary antibodies (Supplementary Table S3) for 16–24 h at 4 °C after 1 h at room temperature (RT) with 5% skim milk, and washed three times at RT for 15 min each. Subsequently, the cells were incubated for 1 h at RT using horseradish peroxidase (HRP)-conjugated IgG (Supplementary Table S3) and underwent another round of three washes, each for 15 min at RT. Finally, proteins from the trimmed membranes were scanned separately using a Pierce SuperSignal West Picochemiluminescent substrate (Merck) with the Chem-Doc XRS system (Bio-Rad, CA, USA).

Cell migration and invasion assays

We employed Transwell polycarbonate membrane chambers (CORNING, NY, USA), both with and without Matrigel (BD, San Jose, CA, USA) pre-coating, to conduct invasion and migration assays respectively. The transfected cells were collected and seeded at 5 × 104 cells/well (for migration assay) or 1 × 105 cells/well (for invasion assay) and were re-suspended into serum-free medium. Cells were seeded into the upper Transwell chamber, and 700 µL of medium containing 10% FBS was added to the lower chamber as an inducer. After incubation for 24 h at 37 °C, the migrated or invaded cells were fixed with 4% paraformaldehyde (Servicebio, Wuhan, China) for 40 min and stained with 0.5% crystal violet (Beyotime, Beijing, China) for 8 min, and cells free in the upper chamber were wiped off with cotton swab gently. Finally, images of migratory or invaded cells in five random fields per chamber were captured under a microscope. The migrated or invaded cells were counted by ImageJ software (NIH, Bethesda, USA).

CCK8 assay

Transfected cells were collected and seeded in 96-well plates at 4 × 103 cells/well. Transfected cell viability was measured at 24 h intervals. At each time point, the mixture contained 10 µL CCK-8 solution (Dojindo, Kumamato, Japan) and 90 µL medium containing serum were added to each well and incubated for 1–2 h at 37 °C. Absorbance at 450 nm (OD450 value) was determined using a microplate reader (Thermo Fisher, Waltham, USA).

Clone formation assay

Transfected cells were collected and 2 × 103 cells/well were seeded in 6-well plates and incubated at 37 °C for 7–10 days, with medium changed every 2–4 days. The cells were fixed with 4% paraformaldehyde (Servicebio, Wuhan, China) for 40 min and stained with 0.5% crystal violet (Beyotime, Beijing, China) for 8 min. Cell counting was carried out using Image J software (NIH, Bethesda, USA).

Statistical analysis

GraphPad Prism 8.0 (GraphPad Prism, CA, USA) was employed for statistical analysis. The statistical difference between the two groups of data was adopted by double-tailed Student’s t-test or paired sample t-test, and the statistical difference between multiple groups of data was adopted by one-way ANOVA test. The presented data represent means ± SD or means ± SE, derived from a minimum of three independent experiments. A p-value less than 0.05 was considered as indicative of statistical significance.

Results

hnRNPA1 expression is upregulated in GC and particularly associated with GC distant metastasis

The analysis of 33 pairs of GC tissues and normal tissues samples from The Cancer Genome Atlas (TCGA) database showed significantly higher levels of hnRNPA1 expression in tumor tissues (Fig. 1A). The level of hnRNPA1 expression was positively correlated with distant metastasis in GC (Fig. 1B). In addition, patients with higher hnRNPA1 expression displayed lower survival rate (Fig. 1C, D).

We examined the expression of hnRNPA1 in human GC cells (AGS, HGC27 and MKN45) and normal human gastric mucosal epithelial cells (GES-1) by qRT-PCR and Western blot. Both mRNA and protein expression levels of hnRNPA1 were significantly higher in GC cell lines than in GES-1 cells (Fig. 1E). These data further validated high expression of hnRNPA1 in GC.

Fig. 1 Differential expression and prognostic analysis of hnRNPA1. A Comparative analysis of hnRNPA1 expression in 33 pairs of GC tissues and adjacent normal tissues, based on data from the TCGA database. B Analysis of hnRNPA1 expression in gastric cancer with and without distant metastasis using GSE26901 database. C Analysis of OS of GC patients through TCGA database using Kaplan-Meier Plotter. D Analysis of PPS of GC patients through TCGA database using Kaplan-Meier Plotter. E hnRNPA1 mRNA and protein table in human normal gastric mucosal epithelial cells (GES-1) and GC cells (AGS, HGC27, and MKN45). *P < 0.05; **P < 0.01; ***P < 0.001

hnRNPA1 promotes GC cells metastasis and proliferation

Tumor progression is manifested by cell metastasis and proliferation. To investigate whether hnRNPA1 would affect the metastatic and proliferative ability of GC cells, we selected AGS cells with the highest hnRNPA1 expression level among the above three GC cells as our experimental cells. Knockdown efficiency after transfection of siRNAs (including sihnRNPA1-1, sihnRNPA1-2 and si-NC as negative control) was determined by qRT-PCR and Western blot. Our findings indicated a substantial reduction in both hnRNPA1 mRNA and protein levels (Fig. 2A). Meanwhile, we examined the overexpression efficiency of the transfected overexpression plasmid (OE-hnRNPA1) and the empty plasmid (pcDNA3.1) by qRT-PCR and Western blot. The results confirmed a significant increase in both hnRNPA1 mRNA and protein levels (Fig. 2B).

Subsequently, we performed cell function assays to analyze the effect of knockdown or overexpression of hnRNPA1 on metastasis and proliferation of AGS cells. The transwell assay demonstrated that the metastatic abilities of the cells were significantly reduced after knockdown of hnRNPA1 (Fig. 2C), whereas the metastasis ability of the cells overexpressing hnRNPA1 was significantly increased (Fig. 2D). Furthermore, in the CCK8 and colony formation assays the cell proliferative ability was significantly reduced after knockdown of hnRNPA1 (Fig. 2E), and overexpressing hnRNPA1 had a significantly increased proliferation capacity (Fig. 2F).

In order to further explore the metastasis of AGS cells, we assessed the expression of EMT-related markers(Snail, Vimentin and E-cadherin)in both the sihnRNPA1 group and si-NC group via Western blot analysis. Our results revealed that knockdown of hnRNPA1 resulted in upregulation of E-cadherin and downregulation of snail and Vimentin, thereby attenuating EMT (Fig. 2G). These findings together suggest that hnRNPA1 promote the metastasis and proliferation of human GC cells, and the process of EMT.

Fig. 2 hnRNPA1 can promote the metastasis and proliferation of GC cells. A The levels of hnRNPA1 mRNA and protein in AGS cells subjected to transfection with sihnRNPA1 or si-NC were quantified using qRT-PCR and Western blot experiments. B The mRNA and protein expression of hnRNPA1 in AGS cells transfected with OE-hnRNPA1 or pcDNA3.1 was determined by qRT-PCR and western blot. C The migration and invasion abilities of AGS cells transfected with sihnRNPA1 or si-NC were assessed by Transwell migration and invasion assay. D The migration and invasion abilities of AGS cells transfected with OE-hnRNPA1 or pcDNA3.1 were determined by Transwell migration and invasion assay. E Proliferation ability in AGS cells of sihnRNPA1 or si-NC was evaluated by CCK-8 assay and colony formation assay. F Proliferation ability in AGS cells of OE-hnRNPA1 or pcDNA3.1 was determined by CCK-8 assay and colony formation assay. G The expression of EMT-related markers in AGS cells transfected with sihnRNPA1 or si-NC was detected by western blot assay. *P < 0.05; **P < 0.01; ***P < 0.001

hnRNPA1 promotes Wnt/β-catenin signaling pathway through WISP2 in GC cells

To further explore the mechanism of hnRNPA1 in AGS cell metastasis and proliferation, expression of Wnt/β-catenin signaling pathway-related markers (GSK-3β, P-GSK-3β and β-catenin) in sihnRNPA1 or OE-hnRNPA1 and their corresponding controls were further examined by Western blot assay. As demonstrated in Fig. 3A, B, knockdown of hnRNPA1 led to up-regulation of GSK-3β and down-regulation of P-GSK-3β and β-catenin, leading to inhibition of Wnt/β-catenin signaling pathway, whereas overexpression of hnRNPA1 led to down-regulation of GSK-3β and up-regulation of P-GSK-3β and β-catenin leading to activation of Wnt/β-catenin signaling pathway. Previous studies have found a positive correlation between the expression of WISP2 and β-catenin in gastric cancer and WISP2 degrading the β-catenin degradation complex. Therefore, we evaluated WISP2 as a possible downstream RNA regulated by hnRNPA1. By qRT-PCR and Western blot, we established that knockdown or overexpression of hnRNPA1 resulted in a correspondingly significant decrease or increase in WISP2 at the mRNA and protein levels (Fig. 3C, D). These results suggest that hnRNPA1 regulate Wnt/β-catenin signaling pathway in GC cells through WISP2.

Fig. 3 hnRNPA1 can regulate Wnt/β-catenin signaling pathway in GC cells through WISP2. A The expression of Wnt/β-catenin signaling pathway-related markers was determined by western blot in AGS cells transfected with sihnRNPA1 or si-NC. B The expression of Wnt/β-catenin signaling pathway-related markers was determined by western blot in AGS cells transfected with OE-hnRNPA1 or pcDNA3.1. C The mRNA and protein expression of WISP2 in AGS cells transfected with sihnRNPA1 or si-NC was determined by qRT-PCR and western blot. D The mRNA and protein expression of WISP2 in AGS cells transfected with OE-hnRNPA1 or pcDNA3.1 was determined by qRT-PCR and western blot. *P < 0.05; **P < 0.01; ***P < 0.001

Knockdown of WISP2 inhibits cell metastasis and proliferation

To further investigate the role of WISP2 in GC cell metastasis and proliferation, we examined the knockdown efficiency after transfection of siRNAs (including siWISP2-1, siWISP2-2 and si-NC as negative control) by qRT-PCR and Western blot. The results demonstrated a substantial reduction in both mRNA and protein expression in AGS cells following WISP2 knockdown (Fig. 4A). Transwell assay showed that the metastatic abilities of the cells were significantly reduced after WISP2 knockdown (Fig. 4B). CCK8 and colony formation assays also showed that the cell proliferative ability was significantly reduced after knockdown of WISP2 (Fig. 4C). These results suggest that knockdown of WISP2 has an inhibitory effect on GC cell metastasis and proliferation.

Fig. 4 Knockdown of WISP2 inhibited the metastasis and proliferation of AGS cells. A Detection of WISP2 mRNA and protein levels in AGS cells subjected to transfection with siWISP2 or si-NC by qRT-PCR and western blot experiments. B Evaluation of the migratory and invasive capacities of AGS cells post-transfection with siWISP2 or si-NC, conducted via Transwell migration and invasion assays. C The proliferation ability in AGS cells transfected with siWISP2 or si-NC was detected by CCK-8 assay and colony formation assay. *P < 0.05; **P < 0.01; ***P < 0.001

hnRNPA1 promotes GC cell metastasis and proliferation through WISP2

To further investigate whether hnRNPA1 mediates GC cell metastasis and proliferation via WISP2, we performed co-transfection (sihnRNPA1 or si-NC, OE-WISP2 or pcDNA3.1) in AGS cells to achieve knockdown of hnRNPA1 and overexpression of WISP2. Transwell assay demonstrated that co-transfection could partially reverse the metastasis abilities of hnRNPA1 knockdown cells (Fig. 5A). The CCK8 and colony formation assays showed that co-transfection partially reversed the proliferation ability of hnRNPA1 knockdown cells (Fig. 5B). These results suggest that hnRNPA1 promote GC cell metastasis and proliferation through WISP2.

Fig. 5 hnRNPA1 promotes GC cell metastasis and proliferation through WISP2. A The migration and invasion abilities of AGS cells co-transfected with sihnRNPA1 or si-NC, OE-WISP2 or pcDNA3.1 were detected by Transwell migration and invasion assay. B The proliferation ability of AGS cells co-transfected with sihnRNPA1 or si-NC, OE-WISP2 or pcDNA3.1 was determined by CCK-8 assay and colony formation assay. *P < 0.05; **P < 0.01; ***P < 0.001

Discussion

Despite tremendous advances in scientific medical research and clinical management of solid cancers in recent years, the survival rate in advance stage gastric cancer remains decimal. The main cause of cancer related death is due to malignant cell unregulated proliferation, dissemination or distant metastasis. Thus, identifying genes affecting the progression of gastric cancer and developing targeted therapy against these genes are of great significance in improving the prognosis and outcome of gastric cancer. Accumulating amounts of evidence have shown that hnRNPA1 is highly expressed in a variety of tumors and serves as a biomarker for early tumor progression and prognosis [29–31]. Chen et al. demonstrated that knockdown of lncRNARP11-81H3.2 inhibited GC metastasis and proliferation by reducing the expression of hnRNPA1 [32]. However, the mechanism of hnRNPA1 in promoting GC metastasis and proliferation has not been studied much. Therefore, in the present study we investigated the mechanism of hnRNPA1 and its downstream pathways in GC metastasis and proliferation.

Through TCGA database analysis and experiments we found that hnRNPA1 expression levels were higher in GC tissues and cells compared to corresponding normal tissues and cells. Knockdown of hnRNPA1 inhibited GC metastasis and proliferation, while overexpression of hnRNPA1 promoted GC metastasis and proliferation. In addition, hnRNPA1 has also been shown to promote EMT in lung cancer by regulating the alternative splicing of LAS1L exon 9 [33]. Therefore, we hypothesized that the process by which hnRNPA1 promotes GC metastasis may be involved in EMT. Indeed, we demonstrated that knockdown of hnRNPA1 inhibited the EMT process in AGS cells mediated by upregulation of E-cadherin expression and downregulation of Vimentin and Snail.

A large number of studies have shown that Wnt/β-catenin signaling pathway is crucial for the occurrence and development of gastric cancer [34–36], so we hypothesized that Wnt/β-catenin signaling pathway may be involved in the regulation of hnRNPA1 on the metastasis or proliferation ability of gastric cancer, our results showed that hnRNPA1 activated the Wnt/β-catenin signaling pathway. Previous research has established that WISP2 is implicated in the metastasis and proliferation of a diverse array of tumors [22, 23, 37]. As WISP2 has been shown to degrade β-catenin degradation complex and lead to the accumulation of β-catenin [21], we hypothesized that hnRNPA1 could regulate WISP2, our results showed that knockdown of hnRNPA1 resulted in a significant reduction in WISP2 levels which further resulted in inactivation of Wnt/β-catenin signaling pathway and ultimately β-catenin degradation whereas overexpression of hnRNPA1 had the opposite effects.

We demonstrated that hnRNPA1 promote GC cell metastasis and proliferation. Mechanistically, hnRNPA1 activates Wnt/β-catenin signaling pathway through WISP2 to induce EMT. However, the limitation of this study is that more in-depth exploration is needed to clarify the regulation mode between hnRNPA1 and WISP2, and furthermore, our experimental findings have not been validated through in vivo experiments. The subsequent stage will involve conducting animal experiments to validate and further elucidate the underlying mechanism. In conclusion, our findings demonstrate that hnRNPA1 plays an important role in promoting GC cell metastasis and proliferation by regulating WISP2-guided Wnt/β-catenin signaling pathway, highlighting its potential as a therapeutic target for GC.

Supplementary Information

Supplementary Material 1.

Abbreviations

GC Gastric cancer

hnRNPA1 Heterogeneous nuclear ribonucleoprotein A1

WISP2 WNT1 Inducible Signaling Pathway Protein 2

EMT Epithelial–mesenchymal transition

TCGA The Cancer Genome Atlas database

PPS Post progression survival

OS Overall survival

CCK-8 Cell counting Kit-8

Acknowledgements

Thanks to all those who participated in this study.

Author contributions

All authors contributed to the study. Chenyang Jiang: Conceptualization, Data curation, Methodology, Validation and written manuscript. Dengfei Xu: Conceptualization, Data curation, Methodology and Validation. Material preparation, data collection and analysis were also performed by Hao Feng, Zirui Ren, Xiang Li, Yuming Chen. Jifeng Yu: Writing, review and editing. Shundong Cang: Conceptualization, Funding acquisition, Supervision. All authors commented on the manuscript. All authors have reviewed and approved the final manuscript.

Funding

This work was supported by grants from Natural Science Foundation of Henan Province (grant no. 22230042050) and science and technology project of Henan Province (Grant No. 212102310161 and LHGJ20220051).

Data availability

The data generated in this study are available upon request from the corresponding author.

Declarations

Ethics approval and consent to participate

No.

Competing interests

The authors declare no competing interests.

Patient consent for publication

No.

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Chenyang Jiang and Dengfei Xu contributed equally to this work.
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