Table 1.
The anthropometric and laboratory investigations of the polycystic ovary syndrome (PCOS) and control groups.
Fig 1.
Identification of differentially expressed RNAs (DERNAs) in fetal side placental tissue of PCOS and control group.
(A) Volcano plots of DERNAs with log2 (foldchange) as the abscissa and -log10 (P value) as the ordinate. The RNAs that exhibited significant up- or down-regulation in PCOS are symbolically represented by red and blue splashes, respectively (|log2FC| ≥ 1, P < 0.05). Gray splashes represent RNAs whose expression is not significantly altered. (B) Heatmaps for DEmRNAs, DEmiRNAs, and DElncRNAs in PCOS and control group (n = 5 in each group). Each row in the dataset corresponds to a specific RNA molecule, while each column represents a unique sample. The color red is indicative of a heightened level of expression, while the color blue signifies a diminished level of expression.
Fig 2.
Functional enrichment analysis of DEmRNAs in fetal side placental tissue of PCOS and control group.
The scatter plot of enriched top 20 Gene Ontology (GO) terms in (A) biological process (BP), (B) cell components (CC), and (C) molecular function (MF). (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment scatter plot of DEmRNAs in PCOS and control group. The x-axis denotes the rich factor, while the y-axis corresponds to the pathway nomenclature. The dot size corresponds to the gene number, and dot color indicates the P-value. All the significantly enriched KEGG pathways are shown.
Fig 3.
Protein-protein interaction (PPI) network creation and hub gene identification.
(A) Top 10 hub genes obtained by three different algorithms (Degree, MNC, and EPC) using Cytohubba plug-in app. MNC: Maximum Neighborhood Component; EPC: Edge Percolated Component. Darker colors indicate higher rankings. (B) Venn diagram of the hub genes from Degree, MNC, and EPC algorithms. (C) PPI network constructed by Metascape analysis of DEmRNAs. (D) Seven sub-network modules were aggregated and extracted from PPI network.
Fig 4.
Landscape of immune cell infiltration in fetal side placental tissue of PCOS and control group.
(A) Bar plot of the proportion of 22 infiltrated immune cells between PCOS and control group. Each color represents a type of immune cell. (B) The violin plot depicted the differences in immune infiltrating between PCOS and normal samples (n = 5 in each group). (C) Heatmap of correlations among 22 types of immune cells. (D) Correlation analysis between eight hub genes and 22 infiltrated immune cells. The colors blue and red are employed to symbolize positive and negative correlation, respectively. The darker the color, the greater the connection. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 5.
KEGG enrichment analysis of the DElncRNAs’ potential cis target transcripts.
Top 20 significantly enriched KEGG pathways of DEmRNAs in PCOS and control group. The x-axis denotes the rich factor, while the y-axis corresponds to the pathway nomenclature. The dot size corresponds to the gene number, and dot color indicates the P-value.
Fig 6.
Creation of the competitive endogenous RNA (ceRNA) regulatory network.
(A) The ceRNA regulatory network included 18 DEmRNAs, 13 DEmiRNAs, and 29 DElncRNAs in PCOS and control group. DEmRNAs, DEmiRNAs, and DElncRNAs are denoted as green triangles, pink circles, and blue diamonds, respectively. (B) A Sankey diagram of the ceRNA network associated with the immune system. The miRNAs interacting with them are in the middle. Each ceRNA network is represented by the same color.
Fig 7.
The validation of DERNAs of the ceRNA network related to the immune system in PCOS and control group using qRT-PCR.
The quantification of mRNA, lncRNA, and miRNA expression levels was conducted utilizing the 2- ΔΔCT approach. ACTB was used as reference genes for mRNA and lncRNA, and U6 was used as reference genes for miRNA. Data were reported as means ± SEM; n = 30 in each group. Two-tailed student’s t-tests were used, and significant differences were considered when the P-value < 0.05. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.