Effects of human microRNAs 100-5p, 192-5p, and 574-3p on proliferation, migration, and gene expression of human immortalized nasopharyngeal cells

Author(s)

Bárbara Grasiele Müller Coan

• Document type: Doctoral Thesis
• Press: Botucatu. 2022-03-23.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Medicina. Botucatu
• Defense date: 2022-02-21
• Advisor: Deilson Elgui de Oliveira; Danillo Pinhal

Abstract

Cancers are malignant neoplastic diseases with complex etiopathogenesis. Transformed cells acquire diverse biological properties that induce alteration in the migration, invasion, and cell survival rates, favoring metastasis formation. This process is under the influence of many factors, including miRNAs. MiRNAs are molecules that canonically prevent mRNA translation, influencing many cellular processes, carcinogenesis and progression of cancers, like nasopharyngeal carcinoma (NPC). NPC is an aggressive cancer with high invasion rates to adjacent tissues, such as the base of the cranium, face sinuses, and orbits, in addition to a high metastatic rate to local lymph nodes. The nonkeratinizing NPC subtype comprehends over 95% of total cases and, when undifferentiated, is highly related to Epstein-Barr virus (EBV) infection. In a recent study of our group, we observed that immortalized nasopharyngeal cells NP69SV40T transfected with the EBV oncoprotein, Latent membrane protein 1 (LMP1), from M81 strain showed increased expression of human miR-100-5p, miR-192-5p, and miR-574-3p compared to NP69SV40T cells transfected with LMP1 from EBV strain B95.8A. Thus, this study aimed to assess the putative effects of each of the identified microRNAs on the viability, migration, and proliferation of human immortalized nasopharyngeal cells (NP69SV40T) cultivated in vitro. Therefore, target prediction and pathway enrichment analysis were performed to infer which pathways could be modulated by each miRNA followed by expression analysis of selected predicted target genes. Additionally, NP69SV40T cells were transfected with miR-100-5p, miR-192-5p, or miR-574-3p mimic to analyze cell behavior. After their transfection with miR-192-5p mimic, we observed a 40% decrease in RAB2A transcript level and a 43% reduction in cell migration. During the pathway enrichment analysis, miR-192-5p showed predicted involvement in pathways such as MAPK, GPCRs, TKR, and TGF-β. NP69SV40T cells transfected with miR-100-5p or miR574-3p reduced transcriptional levels of its targets FZD8 and STC1, respectively. However, in the performed cellular assays, they did not alter cell behavior. The results demonstrate that miR-192-5p acts as a tumor suppressor, answering part of its function in nasopharyngeal cells. This shows its importance in the NPC context and the necessity of further studies of this miRNA. (AU)

• FAPESP's process: 17/22312-5 - Effects of human endogenous microRNAs on the repertoire of cancer stem cells and Cancer progression using Zebrafish as in vivo model
• Grantee: Barbara Grasiele Muller Coan
• Support Opportunities: Scholarships in Brazil - Doctorate