SPP1 promotes anoikis resistance and metastatic phenotypes in hepatocellular carcinoma through PI3K-sensitive signaling
5 affiliations
- 1Department of Blood Transfusion, The First Affiliated Hospital of Ningbo University
- 2Department of Hepatobiliary Surgery, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital
- 3Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Ningbo University
- 4Department of Clinical Laboratory, Jinan Second People’s Hospital
- 5Institute of Pharmacy (Institute of Traditional Chinese Medicine (TCM) Health Industrial Technology), Shandong University of Traditional Chinese Medicine
Abstract
Background: Anoikis resistance enables tumor cells detached from the extracellular matrix to survive during metastatic dissemination, yet its relationship with the immune microenvironment of hepatocellular carcinoma (HCC) remains incompletely understood. Secreted phosphoprotein 1 (SPP1), which encodes osteopontin (OPN), has been implicated in HCC progression. This study investigated the role of SPP1 in anoikis resistance and metastatic phenotypes and evaluated the clinical relevance of circulating OPN in HCC.
Methods: SPP1 was prioritized through integrated analyses of TCGA-LIHC data, including differential expression, survival association, intersection with anoikis-related genes, and pathway enrichment, followed by characterization of the associated tumor microenvironment. Single-cell transcriptomic analysis was performed to determine the cellular distribution of SPP1 and infer potential intercellular communication. SPP1 expression was validated in HCC tissues and cell lines. Gain- and loss-of-function experiments assessed cell viability, clonogenic growth, migration, invasion, and apoptosis under poly-HEMA suspension. The involvement of PI3K signaling was examined using LY294002. Subcutaneous xenograft and experimental pulmonary colonization models were used to evaluate in vivo phenotypes. Serum OPN and alpha-fetoprotein (AFP) were measured in 58 patients with HCC and 36 healthy controls.
Results: SPP1 was upregulated in HCC and associated with shorter overall survival. Tumors with high SPP1 expression exhibited a myeloid- and stroma-enriched microenvironmental profile. Single-cell analysis identified prominent SPP1 expression in myeloid populations and inferred potential SPP1-mediated communication between macrophage subsets and malignant hepatocytes. Increased SPP1 expression was independently confirmed in HCC tissues and cell lines. SPP1 overexpression enhanced cell viability, clonogenic growth, migration, invasion, and resistance to suspension-induced apoptosis, whereas SPP1 silencing produced reciprocal effects. LY294002 attenuated SPP1-induced AKT activation, anoikis resistance, motility, and mesenchymal-marker expression, supporting the involvement of PI3K-sensitive signaling. Bidirectional manipulation of SPP1 produced reciprocal effects on xenograft growth and pulmonary colonization-related phenotypes. Serum OPN concentrations were elevated in HCC and were higher in patients with advanced-stage or metastatic disease. High OPN was associated with shorter overall survival (HR, 1.93; 95% CI, 1.01–3.69) and disease-free survival (HR, 2.29; 95% CI, 1.19–4.39).In the same cohort, the areas under the receiver operating characteristic curve for distinguishing patients with HCC from healthy controls were 0.819 for OPN, 0.781 for AFP, and 0.879 for their combination. These findings represent exploratory same-cohort discrimination and require validation in an independent, clinically representative at-risk population.
Conclusion: SPP1 contributes to anoikis resistance and metastatic phenotypes in HCC, and these phenotypes are sensitive to PI3K inhibition. Single-cell analyses place SPP1 within a myeloid-associated tumor microenvironment, while the inferred communication signals require further experimental validation. Circulating OPN was associated with aggressive HCC features in the present cohort, and its combination with AFP warrants exploratory evaluation in independent, clinically representative at-risk populations.
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BibTeX
@article{zhang2026spp1,
title = {{SPP1 promotes anoikis resistance and metastatic phenotypes in hepatocellular carcinoma through PI3K-sensitive signaling}},
author = {Xinyue Zhang and Honglong Wei and Reguang Pan and Qiming Ying and YanJin Dong and Mingfeng Xia},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1980175},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1980175/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - SPP1 promotes anoikis resistance and metastatic phenotypes in hepatocellular carcinoma through PI3K-sensitive signaling
AU - Xinyue Zhang
AU - Honglong Wei
AU - Reguang Pan
AU - Qiming Ying
AU - YanJin Dong
AU - Mingfeng Xia
PY - 2026
DA - 2026/10/20
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1980175
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1980175/full
AB - Background: Anoikis resistance enables tumor cells detached from the extracellular matrix to survive during metastatic dissemination, yet its relationship with the immune microenvironment of hepatocellular carcinoma (HCC) remains incompletely understood. Secreted phosphoprotein 1 (SPP1), which encodes osteopontin (OPN), has been implicated in HCC progression. This study investigated the role of SPP1 in anoikis resistance and metastatic phenotypes and evaluated the clinical relevance of circulating OPN in HCC. Methods: SPP1 was prioritized through integrated analyses of TCGA-LIHC data, including differential expression, survival association, intersection with anoikis-related genes, and pathway enrichment, followed by characterization of the associated tumor microenvironment. Single-cell transcriptomic analysis was performed to determine the cellular distribution of SPP1 and infer potential intercellular communication. SPP1 expression was validated in HCC tissues and cell lines. Gain- and loss-of-function experiments assessed cell viability, clonogenic growth, migration, invasion, and apoptosis under poly-HEMA suspension. The involvement of PI3K signaling was examined using LY294002. Subcutaneous xenograft and experimental pulmonary colonization models were used to evaluate in vivo phenotypes. Serum OPN and alpha-fetoprotein (AFP) were measured in 58 patients with HCC and 36 healthy controls. Results: SPP1 was upregulated in HCC and associated with shorter overall survival. Tumors with high SPP1 expression exhibited a myeloid- and stroma-enriched microenvironmental profile. Single-cell analysis identified prominent SPP1 expression in myeloid populations and inferred potential SPP1-mediated communication between macrophage subsets and malignant hepatocytes. Increased SPP1 expression was independently confirmed in HCC tissues and cell lines. SPP1 overexpression enhanced cell viability, clonogenic growth, migration, invasion, and resistance to suspension-induced apoptosis, whereas SPP1 silencing produced reciprocal effects. LY294002 attenuated SPP1-induced AKT activation, anoikis resistance, motility, and mesenchymal-marker expression, supporting the involvement of PI3K-sensitive signaling. Bidirectional manipulation of SPP1 produced reciprocal effects on xenograft growth and pulmonary colonization-related phenotypes. Serum OPN concentrations were elevated in HCC and were higher in patients with advanced-stage or metastatic disease. High OPN was associated with shorter overall survival (HR, 1.93; 95% CI, 1.01–3.69) and disease-free survival (HR, 2.29; 95% CI, 1.19–4.39).In the same cohort, the areas under the receiver operating characteristic curve for distinguishing patients with HCC from healthy controls were 0.819 for OPN, 0.781 for AFP, and 0.879 for their combination. These findings represent exploratory same-cohort discrimination and require validation in an independent, clinically representative at-risk population. Conclusion: SPP1 contributes to anoikis resistance and metastatic phenotypes in HCC, and these phenotypes are sensitive to PI3K inhibition. Single-cell analyses place SPP1 within a myeloid-associated tumor microenvironment, while the inferred communication signals require further experimental validation. Circulating OPN was associated with aggressive HCC features in the present cohort, and its combination with AFP warrants exploratory evaluation in independent, clinically representative at-risk populations.
SN - 1664-3224
ER -