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| miRNA let-7a inhibits ACVR1B expression |
| HONG Zipu1, GUAN Kaiyu2, WANG Shao2, XU Shengnan2, HAO Yanyan3, CHEN Xiaoming1 |
| 1.Department of Pediatric Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 2.Institute of Mental Health, the Affiliated Kangning Hospital of Wenzhou Medical University, Wenzhou 325000, China; 3.Department of Clinical Laboratory, Wenzhou Hospital of Traditional Chinese Medicine, Wenzhou 325000, China |
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| Cite this article: |
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HONG Zipu,GUAN Kaiyu,WANG Shao, et al. miRNA let-7a inhibits ACVR1B expression[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2020, 50(1): 7-11.
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Abstract Objective: To investigate the effect of miRNA let-7a on ACVR1B expression. Methods: Bioinformatics analysis predicted that activin receptor type 1B (ACVR1B) is a potential target gene for let-7a. By constructing ACVR1B 3’UTR wild-type and ACVR1B 3’UTR mutant dual-luciferase reporter vector (pmirGLO-ACVR1B 3’UTR-WT and pmirGLO-ACVR1B 3’UTR-MUT), let-7a mimic or let-7a negative control (let-7a NC) were co-transfected into HEK293T cells respectively and the luciferase activity of each group was detected by dual-luciferase reporter system after the cells were collected. qRT-PCR and Western blot were used respectively to detect the ACVR1B mRNA and protein levels after transfection of let-7a mimics, let-7a inhibitor and let-7a NC respectively. Results: The bioinformatics analysis revealed that the human ACVR1B mRNA 3’UTR region (position 77-83) contains perfect complementarity region with a seed sequence of let-7a. The luciferase activity of the let-7a mimic and pmirGLO-ACVR1B 3’UTR-WT co-transfected group was significantly lower than the let-7a NC and pmirGLO-ACVR1B 3’UTR-WT co-transfected group (P<0.05), while the luciferase activities of pmirGLO-ACVR1B 3’UTR-MUT transfected groups were no statistical significance (P>0.05). In HEK293T cells, mRNA and protein levels of ACVR1B in let-7a mimic transfected group were significantly lower than let-7a NC group and let-7a inhibitor group (P<0.05), and the expression levels decreased by 70% and 50% respectively. Conclusion: ACVR1B was a potential target gene directly regulated by let-7a and let-7a might suppress the expression of ACVR1B gene in HEK293T cells.
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Received: 15 February 2019
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