circEYA1 Functions as a Sponge of miR-582-3p to Suppress Cervical Adenocarcinoma Tumorigenesis via Upregulating CXCL14

Abstract

Circular RNAs (circRNAs) function as efficient microRNA (miRNA) sponges that regulate gene expression in the pathogenesis of many human malignancies. However, their roles in cervical adenocarcinoma remain largely unknown. In this study, we aimed to seek novel circRNAs that regulate cervical adenocarcinoma carcinogenesis and to explore their regulatory mechanisms as well as clinical significance. We identified that 24 circRNAs were differentially expressed in cervical adenocarcinoma tissues by RNA sequencing. Among them, circEYA1 was the most significantly downregulated circRNA in cervical adenocarcinoma. In cervical adenocarcinoma cells, circEYA1 overexpression led to suppression of cell viability and colony formation, promotion of apoptosis, and a decrease of the xenograft tumor growth. The mechanism underlying these observations is that circEYA1 functioned as a sponge of miR-582-3p and abrogated its suppression of CXCL14 expression. Consistently, miR-582-3p inhibition phenocopied the biological effects of circEYA1 overexpression in cervical adenocarcinoma cells. Moreover, miR-582-3p overexpression reversed the suppressive behaviors of circEYA1 in vitro and in vivo. In addition, the expression, correlation, and clinical diagnostic value of cir-cEYA1/miR-582-3p/CXCL14 were confirmed in 198 clinical cervical tissue samples. In summary, our findings highlight a novel tumor suppressive role of circEYA1 in cervical adenocarcinoma tumorigenesis and may provide a potential diagnostic marker and therapeutic target for patients with cervical adenocarcinoma.

Seqpac: A New Framework for small RNA analysis in R using Sequence-Based Counts

“…In our HeLa cell experiment, the main fragment was 61 nt, which indicates a unique fragment given that we had a maximum read length of 75 nt. Even though the methods used in Tong et al (25) and Xu et al (26, 27) were restricted to a maximum read length of 50 nt, we found traces of this fragment in the pile of fragments with unverifiable length of ≥ 50 nt. It must be emphasized, however, that we tried to validate the 5’ ETS rRF in yet another dataset, Snoek et al (62) (SRA accession: PRJNA413777), but here we failed to detect anything in the 5’ EST region.…”

“…Despite only having read lengths of 50 nt to our disposal (see Results 7.1), where 5’ ETS rRF of Peak 1 was 61 nt, we found clear traces of this rRF (Figure 8J). Furthermore, to explore the clinical relevance of this finding we downloaded the Xu et al dataset (26, 27). Here sRNA was extract from confirmed cervical tumors and samples from normal cervix.…”

“…The Xu et al 2020 (26, 27) (SRA accession PRJNA607023; ENA download: https://www.ebi.ac.uk/ena/browser/view/PRJNA607023) dataset, used for validating the 5’ ETS rRF of the 45S pre-rRNA (NR_146144.1) in clinical samples. We only used the 8 fastq files obtained by sRNA size-fractions.…”

“…Figure 8J). Furthermore, to explore the clinical relevance of this finding we downloaded the Xu et al dataset (26,27). Here sRNA was extract from confirmed cervical tumors and samples from normal cervix.…”

Gene structure, differential exon usage, and expression of the testis long intergenic non-protein coding RNA 1016 in humans reveals isoform-specific roles in controlling biological processes

“…To independently evaluate and define the LINC01016 gene structure, RNA-Seq datasets (20-22) were obtained, and splice events were determined based on de novo predictions for LINC01016 in the testis (Fig. 4A) and cervix (Fig.…”