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A new and important relationship between miRNA‐147a and PDPK1 in radiotherapy
Author(s) -
Wang LiJuan,
Li NaNa,
Xu SaiJuan,
Zhang Fan,
Hao MingHua,
Yang XianJun,
Cai XinHua,
Qiu PeiYong,
Ji HongLong,
Xu Ping
Publication year - 2018
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.26523
Subject(s) - microrna , biology , luciferase , gene silencing , untranslated region , gene , three prime untranslated region , plasmid , reporter gene , microbiology and biotechnology , transfection , coding region , pi3k/akt/mtor pathway , gene expression , genetics , messenger rna , signal transduction
It was found that the expression level of miR‐147a was significantly increased and the pathway of PI3K/AKT was dramatically inhibited after radiation. In view of the relationship between miRNA and target genes, we put forward the question, what is the relationship between PI3K/AKT and miR‐147a? In order to find the answer to the question, we used bioinformatics techniques to analyze the relationship between miR‐147 (a or b) and PI3K/AKT signaling pathway. miR‐147a overexpression plasmid and PDPK1 3′UTR luciferase reporter gene plasmid were constructed. Dual luciferase reporter gene system validation experiments were carried out on miR‐147a and PDPK1 relationship. The verification experiments were also carried out. Bioinformatics analysis showed that there is a miR‐147a binding site in the non‐coding region (3′UTR) of PDPK1. In the experimental groups transfected with wild type PDPK1 gene of 3′UTR plasmid, the luciferase activity decreased (or increased) significantly in miR‐147a (or inhibitor) group compared with miR‐NC (or anti‐miR‐NC); There was no significant difference between the miR‐147a group (or inhibitor) and the miR‐NC group (or anti‐miR‐NC) in the transfection of PDPK1‐3′UTR‐Mut gene vector. PDPK1 was a target gene for direct regulation of miR‐147a downstream. Verifying test results showed that the expression of PDPK1 mRNA and protein was reduced after overexpression of miR‐147a, which was up‐regulated after silencing miR‐147a in TC, and V79 cells. These results suggest that miR‐147a could be involved in the regulation of PDPK1 transcription by binding to the target site in PDPK1 mRNA 3′UTR, and then regulated AKT.