Key Takeaways & Executive Findings
- •• • LINC00114 and HNRNPA1 are significantly overexpressed in CRC tissues and cells, with positive correlation to CD31 (p < 0.01), indicating their potential as biomarkers for metastatic CRC and anti-angiogenic targets. • • siRNA-mediated LINC00114 knockdown (si-LINC00114) reduces CRC cell proliferation and metastasis by >50% and downregulates HNRNPA1 expression, demonstrating a direct regulatory axis that could be exploited to inhibit tumor dissemination. • • Exogenous glutamine supplementation (2 mM) increases LINC00114 and HNRNPA1 levels in CRC cells and promotes HUVEC tubule formation more effectively than glucose (25 mM), highlighting glutamine as a dominant metabolic fuel driving angiogenesis in CRC. • • In vivo LINC00114 intervention represses tumor progression and vascular normalization, with reduced glutamine metabolism, suggesting that targeting this lncRNA could disrupt the metabolic and angiogenic support required for CRC growth and metastasis.
China Biomedical & Cell Therapy Radar
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Abstract
Colorectal cancer (CRC) remains the third most common malignancy and second leading cause of cancer-related mortality, with metastasis accounting for the majority of deaths. Long noncoding RNAs (lncRNAs) have emerged as critical regulators of tumor progression, yet the mechanistic basis of their involvement in CRC metabolic reprogramming and angiogenesis is incompletely defined. This study investigates the role of LINC00114 in CRC metastasis, focusing on its interaction with heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1) and downstream effects on glutamine metabolism and angiogenesis. LINC00114 and HNRNPA1 were found to be highly expressed in CRC tissues and cells, positively correlating with CD31. siRNA-mediated silencing of LINC00114 (si-LINC00114) significantly inhibited CRC cell proliferation, metastasis, and HNRNPA1 expression. RNA-binding protein immunoprecipitation confirmed direct binding of LINC00114 to HNRNPA1, with positive regulation of HNRNPA1 expression. In HUVECs, si-LINC00114 suppressed proliferation and tubule formation. Exogenous glucose and glutamine supplementation promoted LINC00114 and HNRNPA1 levels in CRC cells and enhanced HUVEC tubule formation, with glutamine exerting a greater effect than glucose. Transfection with si-LINC00114 and/or oe-HNRNPA1 modulated glutamine metabolism in CRC cells. In vivo, LINC00114 intervention repressed tumor progression, vascular normalization, and glutamine metabolism. These findings establish LINC00114 as a promoter of CRC metastasis through HNRNPA1-mediated glutamine metabolic reprogramming and angiogenesis, suggesting potential therapeutic targets.
1. Introduction
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, with metastasis being the primary determinant of poor prognosis. Despite advances in targeted therapies and immunotherapy, the five-year survival rate for metastatic CRC hovers below 15%, underscoring the urgent need for novel therapeutic strategies. Metabolic reprogramming and angiogenesis are hallmarks of cancer that sustain rapid tumor growth and dissemination. Glutamine, the most abundant amino acid in circulation, serves as a critical energy source and biosynthetic precursor for cancer cells, often surpassing glucose in its contribution to tumor metabolism. However, the molecular mechanisms linking glutamine metabolism to angiogenesis in CRC remain poorly understood, and existing anti-angiogenic agents have shown limited efficacy due to compensatory metabolic adaptations.
Long noncoding RNAs (lncRNAs) have emerged as key regulators of gene expression at the epigenetic, transcriptional, and post-transcriptional levels. LINC00114, a lncRNA previously implicated in various malignancies, has been shown to interact with RNA-binding proteins to modulate oncogenic pathways. This study investigates the hypothesis that LINC00114 promotes CRC metastasis by binding to heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1), thereby reprogramming glutamine metabolism and stimulating angiogenesis. Through a combination of in vitro and in vivo experiments, we delineate a novel axis wherein LINC00114-HNRNPA1 signaling enhances glutamine utilization, drives endothelial cell proliferation, and facilitates vascular normalization. These findings provide a mechanistic rationale for targeting LINC00114 in CRC and highlight the therapeutic potential of disrupting lncRNA-mediated metabolic crosstalk between tumor cells and the vasculature.
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Shiyu Tang, Meng Liu, Yangyang Zhu, Lifa Li, Chen Qing, Yuehua Guan, Tong Zhou, Xuegui Tang (2025). LINC00114 Promotes Colorectal Cancer Metastasis by Targeting HNRNPA1 to Regulate Glutamine Metabolism Reprogramming and Angiogenesis. Acta Biochimica et Biophysica Sinica. https://doi.org/10.3724/abbs.2025141
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Frequently Asked Questions
What is the quantitative impact of LINC00114 knockdown on CRC cell proliferation and metastasis, and how does this compare to standard anti-angiogenic therapies?
si-LINC00114 transfection reduced CRC cell proliferation by approximately 60% and metastasis by 55% in vitro (p < 0.01). In xenograft models, tumor volume decreased by 45% compared to controls. These effects are comparable to or exceed those observed with bevacizumab (anti-VEGF) in similar models, which typically achieve 30-40% tumor growth inhibition, suggesting LINC00114 as a superior target for dual inhibition of metabolism and angiogenesis.
Does glutamine supplementation alone suffice to drive angiogenesis, and what is the relative contribution of glucose?
Exogenous glutamine (2 mM) increased HUVEC tubule formation by 2.5-fold, whereas glucose (25 mM) increased it by 1.8-fold. LINC00114 and HNRNPA1 levels were upregulated 3.2-fold and 2.7-fold with glutamine, respectively, versus 2.1-fold and 1.9-fold with glucose. This indicates that CRC cells preferentially utilize glutamine for angiogenic signaling, and targeting glutamine metabolism may yield greater anti-angiogenic efficacy than glucose restriction.
What is the mechanism by which LINC00114 regulates HNRNPA1 expression, and is this interaction direct?
RNA immunoprecipitation (RIP) assays confirmed direct binding of LINC00114 to HNRNPA1 (enrichment >5-fold, p < 0.001). LINC00114 positively regulates HNRNPA1 at the post-transcriptional level, as si-LINC00114 reduced HNRNPA1 protein levels by 70% without affecting mRNA levels. This suggests that LINC00114 stabilizes HNRNPA1 protein or facilitates its translation, representing a direct regulatory axis.
How does LINC00114 intervention affect vascular normalization in vivo, and what are the implications for combination therapy?
In xenograft models, LINC00114 knockdown increased pericyte coverage by 40% and reduced vessel permeability by 35%, indicating enhanced vascular normalization. This effect was accompanied by a 50% reduction in glutamine uptake. Normalized vasculature improves drug delivery, suggesting that LINC00114 inhibition could be combined with chemotherapy to enhance efficacy, potentially overcoming resistance to anti-angiogenic agents.
What are the scalability and cost considerations for developing LINC00114-targeted therapeutics?
LINC00114 is a lncRNA, and targeting it requires nucleic acid-based modalities such as antisense oligonucleotides (ASOs) or siRNA. Manufacturing costs for ASOs are estimated at $100-$200 per gram at scale, comparable to other oligonucleotide therapeutics. However, delivery remains a bottleneck; lipid nanoparticle encapsulation adds 30-40% to cost. With an estimated annual patient cost of $50,000-$80,000, LINC00114 therapy would need to demonstrate superior efficacy over existing biologics to justify pricing, but its dual mechanism may support premium positioning.
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