Abstract | BACKGROUND/AIMS: METHODS: An MTg-AMO carrying the antisense sequences targeting miR-106b, miR-27a and miR-30d was constructed (MTg-AMO106b/27a/30d). Protein levels were determined by Western blot analysis, and transcript levels were detected by real-time RT-PCR (qRT-PCR). Insulin resistance was analysed with glucose consumption and glucose uptake assays. RESULTS: We found that the protein level of glucose transporter 4 (GLUT4), Mitogen-activated protein kinase 14 (MAPK 14), Phosphatidylinositol 3-kinase regulatory subunit beta (PI3K regulatory subunit beta) and mRNA level of Slc2a4 (encode GLUT4), Mapk14 (encode MAPK 14) and Pik3r2 (encode PI3K regulatory subunit beta) were all significantly down-regulated in the skeletal muscle of diabetic rats and in insulin-resistant L6 cells. Overexpression of miR-106b, miR-27a and miR-30d in L6 cells decreased glucose consumption and glucose uptake, and reduced the expression of GLUT4, MAPK 14 and PI3K regulatory subunit beta. Conversely, silencing of endogenous miR-106b, miR-27a and miR-30d in insulin-resistant L6 cells enhanced glucose consumption and glucose uptake, and increased the expression of GLUT4, MAPK 14 and PI3K regulatory subunit beta. MTg-AMO106b/27a/30d up-regulated the protein levels of GLUT4, MAPK 14 and PI3K regulatory subunit beta, enhanced glucose consumption and glucose uptake. CONCLUSION: Our data suggested that miR-106b, miR-27a and miR-30d play crucial roles in the regulation of glucose metabolism by targeting the GLUT4 signalling pathway in L6 cells. Moreover, MTg-AMO106b/27a/30d offers more potent effects than regular singular AMOs.
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Authors | Tong Zhou, Xianhong Meng, Hui Che, Nannan Shen, Dan Xiao, Xiaotong Song, Meihua Liang, Xuelian Fu, Jiaming Ju, Yang Li, Chaoqian Xu, Yong Zhang, Lihong Wang |
Journal | Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
(Cell Physiol Biochem)
Vol. 38
Issue 5
Pg. 2063-78
( 2016)
ISSN: 1421-9778 [Electronic] Germany |
PMID | 27165190
(Publication Type: Journal Article)
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Copyright | © 2016 The Author(s) Published by S. Karger AG, Basel. |
Chemical References |
- 3' Untranslated Regions
- Antagomirs
- Glucose Transporter Type 4
- MicroRNAs
- Phosphoinositide-3 Kinase Inhibitors
- RNA, Messenger
- Pik3cb protein, rat
- Mitogen-Activated Protein Kinase 14
- Glucose
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Topics |
- 3' Untranslated Regions
- Animals
- Antagomirs
(metabolism)
- Base Sequence
- Cell Line
- Diabetes Mellitus, Experimental
(chemically induced, metabolism, pathology)
- Down-Regulation
- Glucose
(metabolism)
- Glucose Transporter Type 4
(antagonists & inhibitors, genetics, metabolism)
- Insulin Resistance
- Male
- MicroRNAs
(antagonists & inhibitors, genetics, metabolism)
- Mitogen-Activated Protein Kinase 14
(antagonists & inhibitors, genetics, metabolism)
- Muscle, Skeletal
(metabolism)
- Phosphatidylinositol 3-Kinases
(genetics, metabolism)
- Phosphoinositide-3 Kinase Inhibitors
- RNA, Messenger
(metabolism)
- Rats
- Rats, Wistar
- Sequence Alignment
- Signal Transduction
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