Tianjin Medical Journal ›› 2019, Vol. 47 ›› Issue (10): 1014-1019.doi: 10.11958/20190638

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Prediction and validation of bioinformatic effects of HOTAIR on miR-152 targeting HLA-G

LI Xiao-juan1, ZHOU Zhi-yi1, WANG Jue2, QIAN Yuan2,3,4△   

  1. 1 Department of Obstetrics, 2 Department of Preclinical Diagnosis, Medical Laboratory, the First Affiliated Hospital of Kunming Medical University, Kunming 650032, China; 3 Institute of Experimental Diagnosis, Research Institute of Yunnan Province; 4 Key Laboratory of Laboratory Medicine of Yunnan Province
  • Received:2019-03-08 Revised:2019-09-05 Published:2019-10-15 Online:2019-11-11
  • Contact: Xiao-Juan LI E-mail:liyuwei0300@163.com
  • Supported by:
     

Abstract: Abstract: Objective To predict and validate the targeting regulation of HOX antisense RNA (HOTAIR) on human leukocyte antigen-G (HLA-G) by upregulating miR-152-3p. Methods Bioinformatics websites and software were used to predict the binding sites of miR-152-3p targeting to HOTAIR and HLA-G 3′untranslated region (UTR). The 3′-UTR and mutant sequences of HOTAIR and HLA-G genes containing the binding sites with miR-152-3p were designed and synthesized, and the wild-type and mutant double luciferase reporter gene vectors of HOTAIR and HLA-G were constructed. After two-enzyme digestion electrophoresis and sequencing, HOTAIR and HLA-G wild-type and mutant double luciferase reporter gene vectors were co-transfected HTR-8/SVneo cells with miR-152-3p mimics and microRNA negative sequence control (mimics NC) respectively. Luciferase activity was detected, and the effects of miR-152-3p on the expression of HOTAIR and HLA-G were observed. Results The results of double enzyme digestion electrophoresis and sequencing showed that the size and sequence of wild type and mutant type HOTAIR and HLA-G gene vectors were consistent with the experimental expectations, and the vectors were successfully constructed. After transfection of miR-152- 3p mimics, the activity of HOTAIR and HLA-G wild-type luciferase decreased significantly (P<0.05), but the expression of luciferase in mutant HOTAIR and HLA-G vectors was not significantly inhibited (P>0.05). Conclusion HOTAIR regulates HLA-G targeting by upregulating the expression of miR-152-3p.

Key words: HLA-G antigens, genes, reporter, HOTAIR, miR-152-3p, HTR-8/SVneo, dual fluorescein reporter gene

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