RESUMO
Endometriosis (EMs)-related infertility commonly has decreased endometrial receptivity and normal decidualization is the basis for establishing and maintaining endometrial receptivity. However, the potential molecular regulatory mechanisms of impaired endometrial decidualization in patients with EMs have not been fully clarified. We confirmed the existence of reduced endometrial receptivity in patients with EMs by scanning electron microscopy and quantitative real-time PCR. Here we identified an lncRNA, named BMPR1B-AS1, which is significantly downregulated in eutopic endometrium in EMs patients and plays an essential role in decidual formation. Furthermore, RNA pull-down, mass spectrometry, RNA immunoprecipitation, and rescue analyses revealed that BMPR1B-AS1 positively regulates decidual formation through interaction with the RNA-binding protein insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). Downregulation of IGF2BP2 led to a decreased stability of BMPR1B-AS1 and inhibition of activation of the SMAD1/5/9 pathway, an inhibitory effect which diminished decidualization in human endometrial stromal cells (hESCs) decidualization. In conclusion, our identified a novel regulatory mechanism in which the IGF2BP2-BMPR1B-AS1-SMAD1/5/9 axis plays a key role in the regulation of decidualization, providing insights into the potential link between abnormal decidualization and infertility in patients with EMs, which will be of clinical significance for the management and treatment of infertility in patients with EMs.
Assuntos
Endometriose , RNA Longo não Codificante , Proteínas de Ligação a RNA , Adulto , Feminino , Humanos , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/genética , Decídua/metabolismo , Decídua/patologia , Endometriose/metabolismo , Endometriose/genética , Endometriose/patologia , Endométrio/metabolismo , Endométrio/patologia , Infertilidade Feminina/metabolismo , Infertilidade Feminina/genética , Infertilidade Feminina/patologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Proteínas de Ligação a RNA/metabolismo , Proteínas de Ligação a RNA/genética , Transdução de Sinais , Células Estromais/metabolismo , Proteínas Smad , Adulto JovemRESUMO
Purpose: This study aimed to reveal the molecular differences in granulosa cells (GCs) from patients with endometriosis (EM). Methods: RNA sequencing was performed on GCs from patients with EM-related infertility (n = 3) and controls (n = 3). Differentially expressed long noncoding RNAs [differentially expressed lncRNAs (DELs), |log2 FC|>4, false discovery rate (FDR) <0.05] and genes [differentially expressed genes (DEGs), |log2 FC|>1.4, FDR <0.05] in patients with EM-related infertility and controls were screened. Protein-protein interaction (PPI) networks of the DEGs were constructed. Then, mRNA-miRNA-lncRNA pairs based on DEGs and DELs were constructed by comprehensive bioinformatics analyses. In addition, overlapping genes identified from both the PPI and mRNA-miRNA-lncRNA pairs were selected. Finally, a competing endogenous RNA (ceRNA) network incorporating transcription factors (TFs) was constructed. Results: A total of 25,806 lncRNAs and 19,684 mRNAs were detected, and 7 DELs and 46 DEGs were identified. Five hub genes from the PPI network were also identified. A single overlapping gene, NR4A2, from both the PPI network and mRNA-miRNA-lncRNA pairs was identified. Finally, a ceRNA network incorporating TFs, including one mRNA (NR4A2), one miRNA (hsa-miR-217), three lncRNAs (XIST, MCM3AP-AS1, and C17orf51), and five TFs (SRF, POLR2A, NRF1, MNT, and TCF7L2), was successfully constructed. Conclusions: The proposed ceRNA network and the prediction of TFs in GCs from EM-related infertility revealed differences in GCs from patients with EM. Importantly, the novel TFs, lncRNAs, miRNAs, and mRNAs involved in the ceRNA network might provide new insights into the underlying molecular mechanisms of EM-related infertility.
Assuntos
Endometriose/genética , Células da Granulosa/patologia , RNA Circular/genética , Adulto , Estudos de Casos e Controles , China , Biologia Computacional/métodos , Endometriose/patologia , Feminino , Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes/genética , Células da Granulosa/metabolismo , Humanos , MicroRNAs/genética , Mapas de Interação de Proteínas/genética , RNA Circular/metabolismo , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Análise de Sequência de RNA/métodos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Background: The molecular biological mechanism of tubal factor infertility (TFI) is still unclear. Long noncoding RNAs (lncRNAs) are considered a major part of the competitive endogenous RNA (ceRNA) network and have attracted growing attention. Our study aimed to explore the regulatory mechanisms of lncRNAs associated with TFI and screen potential key genes related to TFI. Materials and Methods: Differentially expressed lncRNAs (DELs) and differentially expressed genes (DEGs) were identified by comparing normal and TFI expression patterns of lncRNAs and mRNAs in eutopic endometrial tissues obtained from 3 normal and 3 TFI patients during implantation. These data were used to develop a protein-protein interaction (PPI) network of DEGs using the STRING online software. The identified DELs and DEGs were then used to construct a ceRNA network, and the Network Analyzer Tool Kit in Cytoscape was used to analyze the ceRNA network topology and stability. Finally, the overlapping genes present in both the ceRNA and PPI networks were selected as the potential key genes related to TFI. Results: Ninety-six DEGs (59 up and 37 down) and 45 DELs (28 up and 17 down) were identified. Thirty-four DEGs were mapped in a PPI network. A ceRNA network, including two lncRNAs (LINC00305 and DLX6-AS1), four microRNAs (hsa-miR-20b-5p, hsa-miR-17-5p, hsa-miR-107, and hsa-miR-24-3p), and four mRNAs (MAP3K3, HMGB3, FAM103A1, and TMEM209), was successfully constructed. Importantly, a potential key gene (TMEM209) related to TFI was identified. Conclusion: The construction of a ceRNA network related to TFI may help elucidate the regulatory mechanism by which genes and lncRNAs function as ceRNA networks. Importantly, TMEM209 may be further evaluated as potential therapeutic targets for TFI.
Assuntos
Ácidos Nucleicos Livres/genética , Infertilidade Feminina/genética , Adulto , Ácidos Nucleicos Livres/metabolismo , China , Tubas Uterinas/metabolismo , Feminino , Expressão Gênica/genética , Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes , Humanos , MicroRNAs/genética , Mapas de Interação de Proteínas/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Transcriptoma/genéticaRESUMO
Aim: To explore the circular RNA (circRNA) profile in cumulus cells from endometriosis-associated infertility patients. Methods: The expression of circRNAs was profiled by high-throughput sequencing. Sanger sequencing was performed to identify the backsplicing site. Six candidate circRNAs and their parental genes were measured in 30 samples by quantitative reverse transcription-polymerase chainreaction (qRT-PCR). Bioinformatics analysis was performed to predict the functions. Results: A total of 55 upregulated and 41 downregulated differentially expressed circRNAs were detected. Kyoto Encyclopedia of Genes and Genomes data indicated that these target genes were mainly involved in cumulus cell growth- and differentiation-related pathways. Hsa_circ_0072391, hsa_circ_0007299 and hsa_circ_0057799 were significantly increased, and hsa_circ_001533 was significantly decreased in endometriosis-associated infertility patients. Conclusion: The differentially expressed circRNAs might be potentially involved in pathophysiology of endometriosis-associated infertility.
Assuntos
Células do Cúmulo , Endometriose/genética , Infertilidade Feminina/genética , RNA Circular , Adulto , Biologia Computacional , Endometriose/complicações , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Infertilidade Feminina/etiologia , Adulto JovemRESUMO
BACKGROUND: Decidualization of ectopic endometrium often leads to the extensive proliferation of local tissue and is easily misdiagnosed as malignant tumors. The study is aimed at constructing a microRNA- (miRNA-) mRNA network underlying decidualized endometriotic cyst stromal cells (ECSCs). METHODS: All data were collected from the Gene Expression Omnibus (GEO) database. Firstly, the differentially expressed genes (DEGs, adj. P-Val < 0.05, | log FC | ≥1) and miRNAs (DEMs, P-Val < 0.05, â£log FC | ≥1) were analyzed by the limma package. Secondly, we predicted the target genes (TGs) of these DEMs through the TargetScan, miRDB, and miRTarBase databases. The overlapping genes between DEGs and TGs were screened out. Thirdly, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) enrichment analyses of the overlapping genes were performed for integrated discovery, visualization, and annotation. Then, the protein-protein interaction (PPI) network of the overlapping genes was conducted by the STRING database. Finally, we combined the PPI network and the miRNA-mRNA pairs to build a miRNA-mRNA network. RESULTS: There are 29 DEMs and 523 DEGs. Fourteen overlapping genes were screened out, and these genes were significantly enriched in metabolism and immunity. What is more, a miRNA-mRNA network, including 14 mRNAs and 9 miRNAs, was successfully constructed. CONCLUSIONS: Taken together, the miRNA-mRNA regulatory networks described in this study may provide new insights in the decidualization of ECSCs, suggesting further investigations in novel pathogenic mechanisms.
