Fund programs: Natural Science Foundation of Ningxia (No. 2024AAC03688)
Authors:Li Jun1, Bu Yafei2, Chen Jie2, Ding Bo1, Wang Lei1
Keywords:long intergenic non-coding RNA02086; overexpression; gastric cancer; macrophages; polarization; proliferation; Wnt/β-catenin pathway;
DOI:10.19405/j.cnki.issn1000-1492.2026.02.002
〔Abstract〕 Objective To investigate the effect and mechanism of long intergenic non-coding RNA02086 (LINC02086) overexpression mediated macrophage polarization on the proliferation, migration and invasion of gastric cancer cells.Methods The expression levels of LINC02086 in the human gastric epithelial cell line GES-1 and human gastric cancer cell lines HCG-27, NCI-N87, and AGS were determined by qRT-PCR. Human acute monocytic leukemia cells (THP-1) were induced to differentiate into M0 macrophages using phorbol 12-myristate 13-acetate (PMA). HGC-27 cells were infected with either LINC02086 overexpression lentivirus (OE-LINC02086) or its negative control lentivirus (Vector), and the culture supernatants were collected as conditioned medium (CM1). M0 macrophages were co-cultured with the infected HGC-27 cells, and the resulting supernatants were designated as conditioned medium 2 (CM2). M0 macrophages were treated with CM1 alone or in combination with Wnt/β-catenin pathway inhibitor IWR-1, forming the Vector+CM1, OE-LINC02086+CM1, and OE- LINC02086+CM1+IWR-1 groups, respectively. Flow cytometry was used to detect mannose receptor C-type 1 (CD206) expression, and qRT-PCR was employed to measure mRNA levels of interleukin-10 (IL⁃10), transforming growth factor-β (TGF⁃β), vascular endothelial growth factor (VEGF), and chemokine ligand 22 (CCL22). Western blot was performed to evaluate protein expression of CD206, VEGF, and key components of the Wnt/β-catenin pathway—Wnt family member 3a (Wnt3a), glycogen synthase kinase-3β (GSK-3β), and β-catenin. HGC-27 cells were treated with CM2 alone or combined with IWR-1, establishing the Vector+CM2, OE-LINC02086+CM2, and OE-LINC02086+CM2+IWR-1 groups. CCK-8 assay was used to evaluate cell proliferation, and Transwell assays were conducted to assess migration and invasion capabilities.Results Compared with GES-1 cells, the expression levels of LINC02086 were upregulated in HCG-27, NCI-N87, and AGS cells (P < 0.05), with the smallest increase observed in HCG-27 cells. Compared with Vector+CM1 group, the level of CD206 and the expression levels of IL⁃10, TGF⁃β, VEGF and CCL22 mRNA in macrophages stimulated by OE-LINC02086+CM1 increased (P<0.05). Meanwhile, the expression levels of Wnt3a and β-catenin proteins in cells increased (P<0.05), and the expression level of GSK-3β protein decreased (P<0.05). However, co-treatment with IWR-1 markedly reversed the promoting effects of LINC02086 overexpression on the expression of M2 polarization markers, including CD206, IL⁃10, and TGF⁃β mRNA, in macrophages (P<0.05), as well as its activation of the Wnt/β-catenin signaling pathway (P<0.05). Compared with Vector+CM2 group, HGC-27 cells infected with OE-LINC02086+CM2 had increased proliferation activity and increased number of migration and invasion cells (P<0.05). However, the combined intervention of IWR-1 significantly reversed the promotion of LINC02086 overexpression on the proliferation, migration and invasion of HGC-27 cells (P<0.05).Conclusion LINC02086 overexpression promotes the proliferation, migration and invasion of gastric cancer cells by activating Wnt/β-catenin pathway to mediate M2 polarization of macrophages.