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Archive > Volume 33 Issue 6 > 2026,33(6):630-640. DOI:10.3872/j.issn.1007-385X.2026.06.005 Prev Next
Basic Research
Galectin-9 mediates type Ⅰ collagen production by cancer-associated fibroblasts to promote the invasion and migration of pancreatic cancer cells
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Abstract:
[Abstract] Objective: To investigate the role of galectin-9 (Gal-9) in promoting invasion and migration of pancreatic cancer cells and to elucidate the underlying mechanisms. Methods: Publicly available single-cell RNA sequencing datasets were analyzed to characterize the expression landscape of immune checkpoint molecules in pancreatic cancer. Western blotting and ELISA were performed to validate the expression and secretion profiles of galectin-9 (Gal-9) in pancreatic cancer cell lines and clinically relevant models. An orthotopic pancreatic tumor model was established in C57BL/6 mice. Immunohistochemistry, immunofluorescence staining, and flow cytometry were employed to evaluate the effects of Gal-9 on tumor cell proliferation, as well as on the infiltration and functional status of intratumoral CD8? T cells. Cancer-associated fibroblast (CAF) were treated with recombinant Gal-9 protein or tumor cell-derived conditioned medium (CM). Changes in type Ⅰ collagen expression in CAF were assessed by Western blotting, RT- qPCR, and immunofluorescence staining. Co-immunoprecipitation, immunofluorescence staining, and Western blotting were conducted to investigate the interaction between Gal-9 and CD44, as well as the activation of the JAK2/STAT3 signaling pathway. A subcutaneous · · 630 [[PAGE_INDEX=13 FILE=08b612ef-59ff-47e1-960e-f599d97bfff7.pdf]] 赵闯闯, 等. 半乳糖凝集素-9介导肿瘤相关成纤维细胞Ⅰ型胶原生成促进胰腺癌细胞侵袭与迁移xenograft model was established in nude mice by co-implanting Gal-9-overexpressing pancreatic cancer cells with CAF. Sirius Red staining and Masson's trichrome staining were performed to evaluate collagen deposition in tumor tissues. Extracellular matrix (ECM) proteins deposited by CAF were isolated using a decellularization approach. Wound healing assays, Transwell migration assays, and invasion assays were conducted to assess the effects of CAF-derived type Ⅰ collagen on the migratory and invasive capacities of pancreatic cancer cells. Functional rescue experiments were performed using siRNA-mediated knockdown of Gal-9. Results: Compared with classical immune checkpoint molecules PD-L1 and CTLA-4, Galectin-9 (Gal-9) was significantly upregulated in pancreatic cancer (P < 0.05). In vivo experiments demonstrated that Gal-9 promoted tumor growth (P < 0.01), accompanied by increased Ki-67 expression. Meanwhile, Gal-9 markedly reduced intratumoral CD8? T-cell infiltration and increased the proportion of TIM-3? cells, while the expression levels of Granzyme B, IFN-γ, and TNF-α were significantly decreased (P < 0.05). In vitro studies revealed that both exogenous Gal-9 and tumor cell-derived conditioned medium (CM) significantly upregulated type Ⅰ collagen expression in CAF (P < 0.001). Mechanistically, CD44 was detected in the immunoprecipitated complexes using anti-Gal-9 antibodies in CAF, and the protein expression levels of JAK2/STAT3 were correspondingly increased, indicating activation of the JAK2/STAT3 signaling pathway. Functional assays showed that treatment of pancreatic cancer cells with CAF-derived type Ⅰ collagen significantly enhanced their migratory and invasive capacities. In contrast, knockdown of Gal-9 in tumor cells resulted in reduced levels of CAF- derived type Ⅰ collagen, accompanied by a marked decrease in tumor cell migration and invasion. Conclusion: Tumor-derived Gal-9 may exert dual pro-tumorigenic functions in pancreatic cancer. It promotes tumor progression by enhancing cancer cell proliferation and inducing CD8? T cell exhaustion. In parallel, Gal-9 binds to CD44 on CAF to activate the JAK2/STAT3 pathway, thereby stimulating type Ⅰ collagen production and secretion, which in turn facilitates pancreatic cancer cell invasion and migration.
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