MicroRNA-17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis

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MicroRNA-17 expression is increased in adenomyosis tissues and promotes disease by downregulating PTEN, leading to decreased apoptosis and increased cell viability.

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This study evaluated microRNA-17 (miR-17) expression in endometrial tissues from 45 adenomyosis (AM) patients and 32 healthy controls, using RT-qPCR, and investigated miR-17’s function in AM pathogenesis. The authors predicted miR-17 targets and found PTEN mRNA and protein were lower in AM tissues and that antagomiR-17–mediated miR-17 downregulation in cultured endometrial cells increased PTEN expression, decreased Bcl-2, increased Bax, and reduced cyclins E1 and D1, with effects on cell viability and apoptosis assessed by MTT and flow cytometry. PTEN overexpression or miR-17 downregulation both reduced viability and increased apoptosis (P<0.05), and a dual-luciferase reporter assay showed miR-17 directly binds PTEN 3′-UTR to regulate its expression. The paper’s main limitation, as stated by design, is its reliance on in vitro endometrial cell models rather than in vivo functional evidence. This paper is centrally about adenomyosis — specifically miR-17’s PTEN-targeted regulation and downstream effects on apoptosis and cyclin expression in adenomyosis.

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Abstract

The aim of the current study was to evaluate the expression of microRNA (miR)-17 in the endometrial tissues of patients with adenomyosis (AM) and determine its biological function in the occurrence and development of the disease. A total of 45 fresh endometrial tissues of AM patients and 32 normal endometrial tissues were collected from healthy controls. The expression of miR-17 was evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The miR-17-targeting gene phosphatase and tensin homolog (PTEN) was predicted using bioinformatics and its expression was evaluated with RT-qPCR and western blot analysis. Endometrial cells were isolated from patients with AM and healthy controls. They were cultured in vitro and transfected with antagomiR-17 to downregulate miR-17 expression, subsequently cell viability and apoptosis were measured using MTT and flow cytometry. The expression of PTEN and cell cycle- and apoptosis-related proteins were evaluated using western blot analysis. Endometrial cells that stably overexpressed PTEN were screened in vitro by co-culture with G418. A dual-luciferase reporter assay was conducted to verify whether miR-17 was directly bound to PTEN mRNA. The results demonstrated that expression of miR-17 was significantly increased in the endometrial tissues of patients with AM compared with control patients (P<0.05). PTEN mRNA and protein expression were significantly lower in the AM group compared with the control group (P<0.05). When the expression of miR-17 in the cells was downregulated, the expression of PTEN was significantly increased (P<0.05). In addition, expression of Bcl-2 protein was significantly decreased and that of Bax protein significantly increased compared with the negative control (both P<0.05). The expression of cyclins E1 and D1 were also significantly downregulated (P<0.05). When PTEN was overexpressed or miR-17 was downregulated, the viability of endometrial cells significantly decreased and cell apoptosis significantly increased (all P<0.05). A dual-luciferase reporter assay indicated that miR-17 could directly bind to the PTEN mRNA 3'-untranslated region to regulate its expression. Thus the current study indicates that expression of miR-17 was increased in the endometrial tissues of patients with AM and may influence cell apoptosis and cyclin expression through the targeted regulation of PTEN. These results suggest that miR-17 promotes the occurrence and development of AM.
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Print ISSN: 1792-0981 Online ISSN: 1792-1015 International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease. International Journal of Oncology is an international journal devoted to oncology research and cancer treatment. Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery. Oncology Reports is an international journal devoted to fundamental and applied research in Oncology. Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine. Oncology Letters is an international journal devoted to Experimental and Clinical Oncology. Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology. International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis. Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology. Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition. Publishes open-access research on using epigenetics to advance understanding and treatment of human disease. An International Open Access Journal Devoted to General Medicine. Article - Authors: - Pages: 3805-3811|Published online on: August 24, 2017https://doi.org/10.3892/etm.2017.5013 - Expand metrics + The aim of the current study was to evaluate the expression of microRNA (miR)‑17 in the endometrial tissues of patients with adenomyosis (AM) and determine its biological function in the occurrence and development of the disease. A total of 45 fresh endometrial tissues of AM patients and 32 normal endometrial tissues were collected from healthy controls. The expression of miR‑17 was evaluated using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). The miR‑17‑targeting gene phosphatase and tensin homolog (PTEN) was predicted using bioinformatics and its expression was evaluated with RT‑qPCR and western blot analysis. Endometrial cells were isolated from patients with AM and healthy controls. They were cultured in vitro and transfected with antagomiR‑17 to downregulate miR‑17 expression, subsequently cell viability and apoptosis were measured using MTT and flow cytometry. The expression of PTEN and cell cycle‑ and apoptosis‑related proteins were evaluated using western blot analysis. Endometrial cells that stably overexpressed PTEN were screened in vitro by co‑culture with G418. A dual‑luciferase reporter assay was conducted to verify whether miR‑17 was directly bound to PTEN mRNA. The results demonstrated that expression of miR‑17 was significantly increased in the endometrial tissues of patients with AM compared with control patients (P<0.05). PTEN mRNA and protein expression were significantly lower in the AM group compared with the control group (P<0.05). When the expression of miR‑17 in the cells was downregulated, the expression of PTEN was significantly increased (P<0.05). In addition, expression of Bcl‑2 protein was significantly decreased and that of Bax protein significantly increased compared with the negative control (both P<0.05). The expression of cyclins E1 and D1 were also significantly downregulated (P<0.05). When PTEN was overexpressed or miR‑17 was downregulated, the viability of endometrial cells significantly decreased and cell apoptosis significantly increased (all P<0.05). A dual‑luciferase reporter assay indicated that miR‑17 could directly bind to the PTEN mRNA 3'‑untranslated region to regulate its expression. Thus the current study indicates that expression of miR‑17 was increased in the endometrial tissues of patients with AM and may influence cell apoptosis and cyclin expression through the targeted regulation of PTEN. These results suggest that miR‑17 promotes the occurrence and development of AM. Copy and paste a formatted citation Spandidos Publications style Hu H, Li H and He Y: MicroRNA‑17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis. Exp Ther Med 14: 3805-3811, 2017. APA Hu, H., Li, H., & He, Y. (2017). MicroRNA‑17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis. Experimental and Therapeutic Medicine, 14, 3805-3811. https://doi.org/10.3892/etm.2017.5013 MLA Hu, H., Li, H., He, Y."MicroRNA‑17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis". Experimental and Therapeutic Medicine 14.4 (2017): 3805-3811. Chicago Hu, H., Li, H., He, Y."MicroRNA‑17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis". Experimental and Therapeutic Medicine 14, no. 4 (2017): 3805-3811. https://doi.org/10.3892/etm.2017.5013

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nucleotide nitrogen phenol chloroform chlorophenylmethane carbonyl imide fluoride water dimethyl sulfoxide propidium iodide
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rabbits human horseradish naine d'afrique de l'ouest transgenic mice renilla

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