Screening and validation of miRNAs differentially expressed by exosomes of tumor-associated macrophages in microenvironment of salivary gland adenoid cystic carcinoma

doi:10.19438/j.cjoms.2024.03.001

Abstract

Abstract: PURPOSE: To investigate the expression patterns of exosomal miRNAs in tumor-associated macrophages (TAMs) and analyze the potential function in progression of salivary adenoid cystic carcinoma (SACC). METHODS: SACC cells and macrophages were co-cultured to obtain TAMs. Exosomes of both macrophages and TAMs were isolated according to the ultracentrifugation protocol, and then the exosomes were identified using transmission electron microscope, Western blot, and nanoparticle tracking analysis(NTA). RNA-seq analysis was performed to compare the differential expression of miRNAs in TAMs-derived exosomes and the control macrophages-derived exosomes. The target genes of the differential miRNAs were predicted by miRanda and RNAhybrid database. Then GO and KEGG enrichment analysis were performed on the set of target genes. Assays of qRT-PCR, CCK-8, Wound healing, and Transwell were performed to validate the expression patterns and functions of TAMs-derived exosomes. SPSS 22.0 software package was used for data analysis. RESULTS: A total of 1 595 differentially expressed miRNAs were screened out, among which 15 were significantly expressed(P<0.05). Enrichment analysis showed that the target genes of TAMs-derived exosomes were mainly involved in the regulation of cancer-related signaling pathways. Results of qRT-PCR showed that TAMs-derived exosomes carried higher levels of hsa-miR-21-5p than control macrophages derived exosomes. Results of CCK-8, Wound healing, and Transwell assay showed that TAMs-derived exosomal hsa-miR-21-5p promoted proliferation, motility, migration, and invasion of SACC cells. CONCLUSIONS: TAMs-derived exosomal hsa-miR-21-5p promoted malignant progression of SACC cells. TAMs-derived exosomal miRNAs may play important roles in the progression of SACC, and may provide a potential strategy for the diagnosis and treatment of SACC.

Key words: Salivary gland, Adenoid cystic carcinoma, Tumor-associated macrophages, Exosomes, miRNAs, hsa-miR-21-5p

CLC Number:

R739.8

XI Qi, GAO Wan-peng, LI Huan, ZHAO Qi, WANG Jun, WEI Jian-hua, YANG Xin-jie, YANG Zi-hui. Screening and validation of miRNAs differentially expressed by exosomes of tumor-associated macrophages in microenvironment of salivary gland adenoid cystic carcinoma[J]. China Journal of Oral and Maxillofacial Surgery, 2024, 22(3): 209-215.

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