Circular RNA sequencing reveals the molecular mechanism of the effects of acupuncture and moxibustion on endometrial receptivity in patients undergoing infertility treatment

Abstract

The present study aimed to determine the profile of differentially expressed circular RNAs (circRNAs) in infertile patients treated with acupuncture and moxibustion and verify the role of acupuncture and moxibustion in altering endometrial receptivity (ER). High-throughput RNA sequencing and bioinformatics analysis of samples from six pairs of patients treated with or without acupuncture and moxibustion were conducted. The reliability of high-throughput RNA sequencing was validated using reverse transcription-quantitative PCR. The most significant circRNA functions and pathways were selected by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A circRNA-miR-mRNA interaction network was constructed to determine the connection between circRNAs, microRNAs (miRs), and mRNAs. High-throughput RNA sequencing identified 2,653 circRNAs. A total of 86 circRNAs was differentially expressed, of which 57 were upregulated and 29 were downregulated, between the acupuncture and moxibustion group and the control group. In the GO analysis, the identified BP terms were chromatin modification, positive regulation of transcription from RNA polymerase II promoter involved in unfolded protein response, oxidative DNA demethylation, regulation of transcription from RNA polymerase II promoter in response to hypoxia, and regulation of smooth muscle cell differentiation. The identified CC terms were nucleoplasm, nucleolus, nucleus, histone acetyltransferase complex, and annulate lamellae. The identified MF terms were methylcytosine dioxygenase activity, chromatin binding, zinc ion binding, histone binding, and protein binding. In the KEGG pathway analysis, the identified pathways were protein processing in endoplasmic reticulum, degradation of aromatic compounds, shigellosis, mTOR signaling pathway, bacterial invasion of epithelial cells, and prostate cancer. Circ-SFMBT2, circ-BACH1, and circ-LPAR1 were significantly upregulated (P<0.05) and associated with numerous miRs and mRNAs. Acupuncture and moxibustion could impact ER by regulating the expression of circRNAs.

The role of N6-methyladenosine (m6A) modification in the regulation of circRNAs

“…Circular RNA (circRNA) is a class of single-stranded covalently closed RNA molecules that was first discovered in pathogens by Sanger et al in 1976 [1]. It is now generally accepted that circRNA is generated by a process named back-splicing [2], and increasing studies have demonstrated that circRNA plays important roles in the occurrence, development and prognosis of various diseases, including tumorigenesis [3][4][5], neurodevelopmental processes [6] autoimmune responses [7], and infertility [8]. However, studies on how circRNA is regulated before exerting specific biological functions are still limited [9].…”

CircGCN1L1 promotes synoviocyte proliferation and chondrocyte apoptosis by targeting miR-330-3p and TNF-α in TMJ osteoarthritis

“…N = 3 (one sample for three replicates). *p < 0.05. d Transfect miR-330-3p mimics or inhibitor into synoviocytes from the control patient at different concentrations (10,20,50, and 100 nM). N = 6 (two different samples for three independent experiments).…”

“…Total RNA was extracted from five TMJOA tissues and five control tissues using Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. The RNA concentration, purity, and integrity of each sample were quantified, and then the RNA library (mRNA and circRNA) was constructed using the methods described by Shen et al 20 . The RNA libraries were then sequenced by Cloud-seq Biotechnology (Shanghai, China).…”

“…This finding indicated a direct binding of miR-330-3p to TNF. Therefore, we transfected different concentrations (10,20,50, and 100 nM) of miR-330-3p mimics/inhibitor and related controls into TMJ synoviocytes to evaluate the relationship between miR-330-3p and TNF-α. Four concentrations were applied, and the results indicate that TNF-α expression decreased accordingly as the concentration of miR-330-3p mimics increased, which is contrary to the outcome of miR-330-3p inhibition (Fig.…”

Identification of Prostate Cancer-Related Circular RNA Through Bioinformatics Analysis

“…The circRNAs identified in this study have not been reported previously. Therefore, we performed a literature review of circRNAs in PCa in the PubMed database, identifying 13 studies ( Xia et al, 2018 ; Shen et al, 2019 ; Song et al, 2019 ; Wang et al, 2019 ; Wu et al, 2019 ; Hu and Guo, 2020 ; Kong et al, 2020 ; Li T. et al, 2020 ; Ou et al, 2020 ; Shan et al, 2020 ; Yan et al, 2020 ; Zheng et al, 2020a , b ) of 18 circRNA–miRNA interactions. The miRNAs identified in this study have previously been shown to play important roles in PCa.…”