Hypoxia-induced miR-210-3p expression in lung adenocarcinoma potentiates tumor development by regulating CCL2 mediated monocyte infiltration

Sep 21, 2026

Uploaded by Aurora

Hypoxia downregulates CCL2 in many cancers, but the molecular mechanism linking hypoxia to monocyte infiltration and tumor progression remains unclear. This study compares noncancerous and lung adenocarcinoma samples for HIF-1α, miR-210-3p, and CCL2, then tests lung adenocarcinoma cell lines and 3D spheroids. HIF-1α stabilization elevates miR-210-3p and suppresses CCL2, impairing monocyte infiltration. miR-210-3p directly binds the CCL2 3′UTR, and miR-210-3p inhibition promotes antitumor M1 recruitment and tumor regression in a xenograft zebraﬁsh model.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages23

Reading Time58 min

AccessFree to download

Introduction

Hypoxia, lung adenocarcinoma, and tumor-associated macrophages

Mechanistic Findings (HIF-1α/miR-210-3p/CCL2)

Effects on monocyte infiltration and macrophage polarization

Therapeutic Implications

miR-210-3p targeting in xenograft model

Abbreviations

Hypoxia-induced miR-210-3p expression in lung adenocarcinoma potentiates tumor development by regulating CCL2 mediated monocyte inﬁltration Leena Arora 1, Debarun Patra 1, Soumyajit Roy 1, Sidhanta Nanda 1 , Navneet Singh2, Anita K. Verma3, Anuradha Chakraborti4 , Suman Dasgupta5 and Durba Pal 1 1 Department of Biomedical Engineering, Indian Institute of Technology Ropar, Punjab, India 2 Department of Pulmonary Medicine, Postgraduate Institute of Medical Education & Research (PGIMER), Chandigarh, India 3 Department of Zoology, Kirori Mal College, University of Delhi, India 4 Department of Experimental Medicine & Biotechnology, Postgraduate Institute of Medical Education & Research (PGIMER), Chandigarh, India 5 Department of Molecular Biology & Biotechnology, Tezpur University, Assam, India Keywords CCL2; HIF-1Α; LUAD; miR-210-3p; monocyte inﬁltration Correspondence D. Pal, Department of Biomedical Engineering, Indian Institute of Technology Ropar, Rupnagar 140001, Punjab, India Tel: +91 01881 23 2506 E-mail: [durba.pal@iitrpr.ac.in](durba.pal@iitrpr.ac.in) (Received 16 December 2021, revised 24 April 2022, accepted 2 June 2022, available online 22 March 2024) doi:10 . 1002/1878-0261 .13260 In most cancers, tumor hypoxia downregulates the expression of C-C motif chemokine 2 (CCL2), and this downregulation has been implicated in monocyte inﬁltration and tumor progression; however, the molecular mechanism is not yet clear. We compared noncancerous and lungadenocarcinoma human samples for hypoxia-inducible factor 1-alpha (HIF-1A), microRNA-210-3p (mir-210-3p), and CCL2 levels. Mechanistic studies were performed on lung adenocarcinoma cell lines and 3D tumor spheroids to understand the role of hypoxia-induced miR-210-3p in the regulation of CCL2 expression and macrophage polarization. HIF-1Α stabilization increases miR-210-3p levels in lung adenocarcinoma and impairs monocyte inﬁltration by inhibiting CCL2 expression. Mechanistically, miR- 210-3p directly binds to the 30untranslated region (UTR) of CCL2 mRNA and silences it. Suppressing miR-210-3p substantially downregulates the effect of hypoxia on CCL2 expression. Monocyte migration is signiﬁcantly hampered in miR-210-3p mimic-transfected HIF-1A silenced cancer cells. In contrast, inhibition of miR-210-3p in HIF-1A-overexpressed cells markedly restored monocyte migration, highlighting a direct link between the miR- 210-3p level and tumor monocyte burden. Moreover, miR-210-3p inhibition in 3D tumor spheroids promotes monocyte recruitment and skewing towards an antitumor M1 phenotype. Anti-hsa-miR-210-3p-locked nucleic acid (LNA) delivery in a lung tumor xenograft zebraﬁsh model caused tumor regression, suggesting that miR-210-3p could be a promising target for immunomodulatory therapeutic strategies against lung adenocarcinoma. Abbreviations 30 UTR, 30 untranslated region; 5-Aza, 5-Aza-20-deoxycytidine; 5mC, 5-methylcytosine; AhR, aryl hydrocarbon receptor; ARNT, AhR nuclear translocator; CCL2, CC chemokine ligand 2; CCR2, C-C chemokine receptor type 2; CD, cluster of differentiation; cDNA, complementary deoxyribonucleic acid; CoCl2, cobalt chloride; DAPI, 40 ,6-diamidino-2-phenylindole; DNMT, DNA N-methyl transferase; ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HIF-1Α, hypoxia inducible factor-1Α; HRE, hypoxia response element; HUVEC, human umbilical vein endothelial cell; Hypoxa miR, hypoxia-associated microRNA; IL, interleukin; LNA, locked nucleic acid; LUAD, lung adenocarcinoma; miR-210-3p, microRNA-210-3p; MPEG1 . 1, macrophage expressed 1-tandem duplicate 1; MPX, myeloid-speciﬁcperoxidase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NCCS, National Centre for Cell Science; NSCLC, non-small-cell lung cancer; OCT, optimal cutting temperature; ODDD, oxygen-dependent death domain; OE, overexpression; PER, period circadian protein; PHD, prolyl hydroxylase domain; pVHL, protein Von

FAQs

How does hypoxia influence miR-210-3p and CCL2 in lung adenocarcinoma?

HIF-1α stabilization under hypoxic conditions increases miR-210-3p levels while inhibiting CCL2 expression. This establishes a functional link between hypoxia and the CCL2-mediated immune recruitment axis.

What is the mechanism by which miR-210-3p regulates CCL2?

miR-210-3p directly binds the 3′UTR of CCL2 mRNA and silences it. Suppressing miR-210-3p substantially reverses the hypoxia-driven changes in CCL2 expression.

What happens to monocyte infiltration and macrophage phenotype when miR-210-3p is inhibited?

Inhibition of miR-210-3p restores monocyte migration in cells with altered HIF-1α activity. In 3D tumor spheroids, miR-210-3p inhibition promotes monocyte recruitment and skewing toward an antitumor M1 phenotype.

Sep 21, 2026

Uploaded by Aurora