GC-MSCs transcriptionally upregulate SALL4 in gastric cancer through miR-4669/TIMP3/β-catenin signaling

  • Cell Signal. 2025 Jun:130:111668. doi: 10.1016/j.cellsig.2025.111668.
Jing Wen  1 Xiaoli Cao  2 Baocheng Zhou  3 Fang Yang  1 Xiang Wang  1 Yuanyuan Li  1 Xinlan Zhao  1 Jingyu Mei  1 Wei Zhu  1 Li Sun  4 Feng Huang  5 Mei Wang  6
Affiliations
  • 1. Department of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu Province 212013, China.
  • 2. Department of Laboratory Medicine, Affiliated Tumor Hospital of Nantong University, Nantong, Jiangsu Province 226321, China.
  • 3. Department of Medical Laboratory, Lianyungang Maternal and Child Health Hospital, Lianyungang, Jiangsu Province 222000, China.
  • 4. Department of Clinical Laboratory, Kunshan First People's Hospital, Affiliated to Jiangsu University, Kunshan 215300, China.
  • 5. Department of Clinical Laboratory, Kunshan First People's Hospital, Affiliated to Jiangsu University, Kunshan 215300, China; Department of Clinical Laboratory, Maternal and Child Health Care Hospital of Kunshan, Suzhou, Jiangsu Province, China.
  • 6. Department of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu Province 212013, China. Electronic address: [email protected].
Abstract

Backgrounds: Gastric cancer-associated mesenchymal stem cells (GC-MSCs) as integral components of the tumor microenvironment potentiate gastric Cancer growth and metastasis. SALL4 is aberrantly upregulated in gastric Cancer and pivotal for malignant progression. Whether GC-MSCs is responsible for SALL4 upregulation and the underlying mechanisms remains elusive.

Methods: Cancer growth and metastasis capacities were assessed by cell colony formation assay, transwell assay, and epithelial-mesenchymal transition protein detection in vitro as well as subcutaneous xenograft and peritoneal metastasis models in vivo. SALL4 was measured by qPCR, western blot and immunohistochemistry staining. Gain- and loss-functional analysis were performed for miRNA and target gene. β-catenin signaling was assessed by immunofluorescence staining and Top/FopFlash luciferase assay. Transcriptional regulation was conducted using chemicals, luciferase reporter and ChIP assay. Clinical tissues and TCGA-STAD database were included for expression profile, correlation and clinical relevance analysis.

Results: GC-MSCs promoted gastric Cancer growth and metastasis along with elevation of SALL4 and miR-4669 in Cancer cells and tissues. Overexpression of miR-4669 mimicked GC-MSC effects, while miR-4669 knockdown eliminated their oncogenic roles. TIMP3 was identified as a target of miR-4669 and mediated its functions. TIMP3 overexpression counteracted GC-MSC-induced Cancer progression and SALL4 expression. GC-MSCs activated SALL4 transcription through the miR-4669/TIMP3/β-catenin pathway. The regulatory axis was aberrantly expressed in gastric Cancer tissues, correlated with each Other in certain Cancer tissues and associated with lymph node metastasis.

Conclusions: GC-MSCs transcriptionally upregulate SALL4 to facilitate gastric Cancer cell growth and metastasis via miR-4669/TIMP3/β-catenin pathway, highlighting the crucial role of GC-MSCs in the aberrant upregulation of SALL4.

Keywords
Gastric cancer; Mesenchymal stem cells; SALL4; Tumor microenvironment; microRNA.
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