ABSTRACT
BACKGROUND: Cervical cancer is the leading cause of cancer deaths among women, and more than 85% of cervical cancer deaths occur in low and middle-income countries. The purpose of this study is to investigate the functions of MAGI2-AS3 and miR-15b in cervical cancer. MATERIALS AND METHODS: The mRNA levels of MAGI2-AS3, miR-15b, and CCNE1 were evaluated using RT-qPCR assay. Dual-luciferase reporter gene assay was used to confirm whether miR-15b binds to CCNE1. RESULTS: LncRNA MAGI2-AS3 was downregulated, while miR-15b was upregulated in cervical cancer. Cervical cancer patients with low expression of MAGI2-AS3 have a poor prognosis. Upregulation of MAGI2-AS3 inhibited proliferative and invasive abilities of HeLa cells via regulating the expression of miRNA-15b. MiR-15b inhibitor suppressed cell proliferation and invasion. CCNE1 was a direct target gene of miR-15b, which binds to the 3'-UTR of its mRNA. MiR-15b partially reversed the inhibitory effect of overexpression of MAGI2-AS3 on the proliferation and invasion of HeLa cells. MAGI2-AS3 mediated the expression of CCNE1 in HeLa cells. CONCLUSION: LncRNA MAGI2-AS3 inhibits the proliferation and invasion of cervical cancer cells via the miRNA-15/CCNE1 axis. Our results illustrates that MAGI2-AS3 can be used as a useful clinical predictor for early diagnosis and prognosis assessment of cervical cancer.
Subject(s)
MicroRNAs , RNA, Long Noncoding , Uterine Cervical Neoplasms , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Cell Line, Tumor , Cell Proliferation , Female , Gene Expression Regulation, Neoplastic , Guanylate Kinases/genetics , Guanylate Kinases/metabolism , HeLa Cells , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , RNA, Messenger , Uterine Cervical Neoplasms/geneticsABSTRACT
MicroRNAs are a group of small non-coding RNAs that play key roles in almost every aspect of mammalian cell. In kidney, microRNAs are required for maintaining normal function of renal cells, disruption of which contributes to pathogenesis of renal diseases. In this study, we investigated the potential role of miRNAs as key regulators of podocyte survival by using a primary cell culture model from non-human primates (NHPs). Through microRNA profile comparison in glomeruli from mouse, rat and NHP, miR-27b was found to be among a list of glomeruli-enriched miRNA conserved across species. In NHP primary podocyte culture, significant downregulation of miR-27b was observed during treatment of puromycin aminonucleoside (PAN), a classic nephrotoxin. Overexpression of miR-27b enhanced PAN-induced apoptosis and cytoskeleton destruction in podocytes while its inhibition had a protective effect. Target identification analysis identified Adora2b as a potential direct target of miR-27b. Ectopic expression of miR-27b suppressed both Adora2b mRNA and protein expression, whereas inhibition of miR-27b increased the transcript and protein expression levels of Adora2B. Dual luciferase assay further confirmed Adora2b as a direct target of miR-27b. Furthermore, knockdown of Adora2b by siRNAs enhanced PAN-induced apoptosis, similar to the phenotypes we had observed with miR-27b overexpression. In addition, stimulating the adenosine signaling by an Adora2b agonist, NECA, improved podocyte survival upon PAN treatment. Taken together, our data identified a novel role of miR-27b-adora2b axis in primary podocyte survival upon injury and suggested a critical role of adenosine signaling pathway in podocyte protection.