miR‐27b‐5p inhibits BeWo cells fusion by regulating WNT2B and enzyme involved in progesterone synthesis
| Autor: | Sonam Verma, Ankita Malik, Piyush Chaudhary, Satish K. Gupta, Richa Mishra, Amulya K. Panda, Sudha Saryu Malhotra |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Placenta Immunology Steroid Isomerases Cell Fusion 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Western blot Multienzyme Complexes Pregnancy Cell Line Tumor microRNA medicine Humans Immunology and Allergy Gene silencing Secretion Progesterone Glycoproteins chemistry.chemical_classification 030219 obstetrics & reproductive medicine Forskolin medicine.diagnostic_test Progesterone Reductase Colforsin Obstetrics and Gynecology Progesterone secretion Transfection Cell biology Wnt Proteins MicroRNAs 030104 developmental biology Enzyme Reproductive Medicine chemistry embryonic structures Female |
| Zdroj: | American Journal of Reproductive Immunology. |
| ISSN: | 1600-0897 1046-7408 |
| DOI: | 10.1111/aji.13409 |
| Popis: | Problem The miRNAs show placenta-specific expression patterns, which alter during pregnancy-related complications. In present study, the role of miR-27b-5p during forskolin-mediated BeWo cells fusion has been investigated. Method of study The fusion of BeWo cells in response to forskolin treatment (25 µM) was studied by desmoplakin I+II staining. Expression profile of miR-27b-5p by qRT-PCR and its targets HSD3β1 and WNT2B by qRT-PCR and in Western blot were studied. The effect of overexpression of miR-27b-5p and silencing of HSD3β1 & WNT2B by siRNA on forskolin-mediated BeWo cells fusion and secretion of hCG and progesterone by ELISA was investigated. Results Time-dependent down-regulation in the expression of miR-27b-5p in forskolin-treated BeWo cells has been confirmed by qRT-PCR. Overexpression of miR-27b-5p significantly inhibits forskolin-mediated BeWo cells fusion as well as hCG & progesterone secretion. HSD3β1 and WNT2B were identified as targets of miR-27b-5p and are up-regulated in forskolin-treated BeWo cells. Overexpression of miR-27b-5p in BeWo cells downregulates their expression. Further, luciferase reporter assay revealed that miR-27b-5p directly target expression of both HSD3β1 and WNT2B. Silencing of both HSD3β1 and WNT2B leads to a significant reduction in forskolin-mediated BeWo cells fusion with concomitant decrease in the secretion of progesterone or/and hCG. Decrease in forskolin-mediated cells fusion observed in miR-27b-5p mimic transfected BeWo cells could be rescued by the overexpression of both HSD3β1 and WNT2B. Conclusion These observations suggest that reduced miR-27b-5p in forskolin-treated BeWo cells leads to increased secretion of progesterone and hCG due to loss of repressional control on HSD3β1 and WNT2B. |
| Databáze: | OpenAIRE |
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