Tohoku J. Exp. Med., 2018 October, 246(2)

Deregulated MicroRNAs in Cancer-Associated Fibroblasts from Front Tumor Tissues of Lung Adenocarcinoma as Potential Predictors of Tumor Promotion

MARÍA CRISTINA NEGRETE-GARCIA,1,2 SANDRA LIZBETH RAMÍREZ-RODRIGUEZ,1 CLAUDIA RANGEL-ESCAREÑO,3 SAID MUÑOZ-MONTERO,3 JAVIER KELLY-GARCÍA,4 MARÍA EUGENIA VÁZQUEZ-MANRÍQUEZ,5 PATRICIO SANTILLÁN,6 MARTHA MONTAÑO RAMÍREZ,7 GUSTAVO RAMÍREZ-MARTÍNEZ,8 ALEJANDRA RAMÍREZ-VENEGAS9 and BLANCA ORTIZ-QUINTERO1

1Research Unit, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico
2Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina del Instituto Politécnico Nacional, Mexico City, Mexico
3Instituto Nacional de Medicina Genómica, Mexico City, Mexico
4Oncology Service, Centro Médico Siglo XXI, Mexico City, Mexico
5Department of Pathology, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico
6Department of Thoracic Surgery, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico
7Department of Cellular Biology, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico
8Department of Immunobiology and Genetics, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico
9Smoking and COPD Research Department, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas,” Mexico City, Mexico

Cancer-associated fibroblasts (CAFs) are the main component of the tumor stroma and promote tumor progression through several mechanisms. Recent evidence indicates that small noncoding RNAs, microRNAs (miRNAs), play key roles in CAF tumor-promoting properties; however, the role of miRNAs in lung cancer-associated fibroblasts remains poorly defined. We characterized the differential miRNA expression profile of fibroblasts isolated from matched tumor front (F-CAFs), inner tumor (In-CAFs), and normal adjacent (NFs) tissues from four lung adenocarcinoma patients (ADs) using microarray analysis. Proliferation and invasion assays of A549 human lung cancer cells in the presence of conditioned medium from F-CAFs, In-CAFs or NFs were performed to assess tumorigenic properties. Ten identified candidate miRNAs in F-CAFs, In-CAFs and NFs from 12 ADs were then validated by RT-PCR. Both F-CAFs and In-CAFs enhanced the proliferation and invasion of A549 cells compared with NFs; moreover, F-CAFs showed a significantly stronger effect than In-CAFs. RT-PCR validation demonstrated three downregulated miRNAs in F-CAFs compared with NFs (miR-145-3p, miR-299-3p, and miR-505-3p), two in F-CAFs compared with In-CAFs (miR-410-3p and miR-485-5p), but no differentially expressed miRNAs between In-CAFs and NFs. Further target-gene prediction and pathway enrichment analysis indicated that deregulated miRNAs in F-CAFs showed significant associations with “pathways in cancer” (miR-145-3p, miR-299-3p and miR-410-3p), “Wnt signaling pathway” (miR-410-3p and miR-505-3p), and “TGF-beta signaling pathway” (miR-410-3p). Importantly, a tumor-promoting growth factor targeted by those miRNAs, VEGFA, was upregulated in F-CAFs compared with NFs, as judged by RT-PCR. In conclusion, deregulated miRNAs in F-CAFs are potentially associated with CAF tumor-promoting properties.

Key words —— cancer-associated fibroblast; lung adenocarcinoma; microenvironment heterogeneity; microRNA; tumor stroma

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Tohoku J. Exp. Med., 2018, 246, 107-120

Correspondence: Blanca Ortiz-Quintero, Ph.D., Research Unit. Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas”. Calzada de Tlalpan 4502, Colonia Sección XVI, Mexico City 14080, Mexico.

e-mail: boq@iner.gob.mx