miR-7/TGF- β2 axis sustains acidic tumor microenvironment-induced lung cancer metastasis

Abstract

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.

Keywords: AOM/DSS, azoxymethane/dextran sodium sulfate; Acidic tumor microenvironment; B[a]P, benzopyrene; CA9, carbonic anhydrase IX; DAB, diaminobenzidine; DAVID, Database for Annotation, Visualization, and Integrated Discovery; DEGs, differentially expressed genes; DEN, diethylnitrosamine; DEPs, differentially expressed proteins; DSS, dextran sodium sulfate; GEMMs, genetically engineered tumor mouse models; GSEA, gene set enrichment analysis; IHC, immunohistochemistry; ISH, in situ hybridization; Invasion; KEGG, Kyoto Encyclopedia of Genes and Genomes; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; Lung cancer; MCT, monocarboxylate transporter; Metastasis; NHE, Na+/H+ exchanger; NSCLC, non-small cell lung cancer; PCR, polymerase chain reaction; TGF-β2; TME, tumor microenvironment; TMT, tandem mass tagging; V-ATPase, vacuolar ATPase; miR-7-5p; pH.