Investigations on the mechanism of progesterone in inhibiting endometrial cancer cell cycle and viability via regulation of long noncoding RNA NEAT1/microRNA-146b-5p mediated Wnt/β-catenin signaling.

Progesterone is often used to protect the endometrium and prevent endometrial cancer. An intensive study on its molecular mechanism in endometrial cancer would contribute to the development of more promising therapies. Relevant lncRNAs and mRNAs expression data in endometrial cancer cell line Ishikawa pretreated and post-treated with progesterone were derived from Gene Expression Omnibus (accession no. GSE29435), and then we analyzed long noncoding RNAs and mRNAs with differential expressions in two different conditions. The Cytoscape software, TargetScan, miRanda, and Human microRNA Disease Database (HMDD) websites were employed. Gene set enrichment analysis (GSEA) was used to determine related Kyoto Encyclopedia of Genes and Genomes pathways alteration in Ishikawa cells treated with progesterone. In addition to bioinformatics analysis, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western blot, and dual-luciferase reporter assays were performed. The impact of progesterone on cell propagation and cell cycle was testified by colony formation and flow cytometry analysis. LncRNA nuclear enriched abundant transcript 1 (NEAT1) was the most significantly downregulated lncRNA in endometrial cancer cells treated with progesterone. Lymphoid enhancing factor 1 (LEF1) was positively associated with NEAT1, and eventually hsa_miR-146b-5p was validated to target both LEF1 and NEAT1. Wnt/β-catenin signaling pathway was identified to involve in endometrial cancer. NEAT1 or LEF1 was overexpressed in endometrial cancer cells while downregulated following post-treatment with progesterone. Conversely, miR-146b-5p was notably decreased in Ishikawa cells while upregulated after treatment with progesterone. Downstream gene c-myc or MMP9 regulated by upstream gene LEF1 in Wnt/β-catenin signaling pathway was remarkably increased in Ishikawa cells and positively related with NEAT1. Progesterone inhibited cell cycle and viability through regulating NEAT1/miR-146b-5p axis via Wnt/β-catenin signaling pathway. Progesterone exerted suppressive influence on endometrial cancer progression via regulation of lncRNA NEAT1/miR-146b-5p-mediated Wnt/β-catenin signaling pathway, which might reveal new strategies for developing more effective therapeutics. © 2018 IUBMB Life, 71(1):223-234, 2019.

Huang X ，Zhong R ，He X ，Deng Q ，Peng X ，Li J ，Luo X ... -  《-》

Regulation of NEAT1/miR-214-3p on the growth, migration and invasion of endometrial carcinoma cells.

Wang J ，Zhao X ，Guo Z ，Ma X ，Song Y ，Guo Y ... -  《-》

The Long Noncoding RNA NEAT1 Targets miR-34a-5p and Drives Nasopharyngeal Carcinoma Progression via Wnt/β-Catenin Signaling.

Long noncoding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) has been deemed an oncogene in many human cancers. However, the underlying mechanism of NEAT1 in nasopharyngeal carcinoma (NPC) progression remains largely unclear. Quantitative real-time PCR assay was performed to assess the expression of NEAT1 and miR-34a-5p in NPC tissues and cells. Western blot analysis was used to observe cell epithelial to mesenchymal transition (EMT) and the activation of Wnt/β-catenin signaling in 5-8F cells. MiRNA directly interacting with NEAT1 were verified by dual-luciferase reporter assay and RNA immunoprecipitation. Cell proliferation ability was determined by CCK-8 assay, and cell migration and invasion capacities were assessed by transwell assays. An animal model was used to investigate the regulatory effect of NEAT1 on tumor growth in vivo. Our data revealed that NEAT1 is upregulated, while miR-34a-5p is downregulated in NPC tissues and cell lines. NEAT1 knockdown repressed tumor growth in vitro and in vivo. Additionally, we discovered that NEAT1 directly binds to miR-34a-5p and suppresses miR-34a-5p expression. Moreover, NEAT1 knockdown exerted suppression effects on cell proliferation, migration, invasion, and EMT by miR-34a-5p. NEAT1 knockdown blocked Wnt/β-catenin signaling via miR-34a-5p. Our study demonstrated that NEAT1 targets miR-34a-5p at least partly to drive NPC progression by regulating Wnt/β-catenin signaling, suggesting a potential therapeutic target for NPC.

Ji Y ，Wang M ，Li X ，Cui F ... -  《-》

Progesterone ameliorates the endometrial polyp by modulating the signaling pathway of Wnt and β-catenin via regulating the expression of H19 and miR-152.

MicroRNAs and long noncoding RNAs are believed to play important roles in the pathogenesis of various diseases. This study aimed to explore the potential mechanism of the involvement of H19 and miR-152 in an endometrial polyp. Luciferase assay was conducted to determine the effect of progesterone. Real-time polymerase chain reaction (PCR) and western blot were performed to detect the influence of progesterone on miR-152 and Wnt1. MTT assay and flow cytometry (FCM) were utilized to detect the effect of progesterone on cell proliferation and apoptosis. In silicon analysis, luciferase assay, real-time PCR, and immunohistochemistry (IHC) were performed to explore the regulatory relationship between H19 and miR-152 or miR-152 and Wnt1. Progesterone dose-dependently increased the H19 expression level through driving the promoter efficiency of H19. Then, progesterone upregulated Wnt1 level and downregulated miR-152 in a dose-dependent manner in ECC1 and HEC1A cells. Administration of progesterone inhibited cell viability and promoted cell apoptosis. H19 negatively regulated miR-152 expression by binding to miR-152. Furthermore, Wnt1 was identified as a virtual target gene of miR-152 and was inhibited by miR-152. Progesterone receptors mRNA and miR-152 were lowly expressed in participants with an endometrial polyp, while the levels of H19 and Wnt1 were much higher in the endometrial polyp group compared with normal controls. H19 negatively regulated miR-152 and miR-152 negatively regulated Wnt1, with the negative correlation coefficients being -0.500 and -0.500, respectively. Using IHC, it was found that Wnt1 and Bcl-2 protein were highly expressed in the endometrial polyp group compared with normal controls. The results suggested that H19 was associated with endometrial polyp via mediating cell proliferation and apoptosis.

Feng M ，Zhang T ，Ma H 《-》

Upregulation lnc-NEAT1 contributes to colorectal cancer progression through sponging miR-486-5p and activating NR4A1/Wnt/β-catenin pathway.

Liu Z ，Gu Y ，Cheng X ，Jiang H ，Huang Y ，Zhang Y ，Yu G ，Cheng Y ，Zhou L ... -  《-》