Abstract
MicroRNA (miR)-214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR-214 and ATF4 were involved in the MEG3-mediated increase of FoxO1 expression. MEG3, miR-214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR-214 and ATF4 was analysed using the luciferase reporter assay. MEG3-targeting small interference RNAs were injected into high-fat diet (HFD)-fed mice to verify the role of MEG3 in hepatic insulin resistance in vivo. MEG-3 and ATF4 were demonstrated to be upregulated and miR-214 was indicated to be downregulated in the livers of HFD-fed and ob/ob mice. In mouse primary hepatocytes, palmitate time-dependently increased MEG3 and ATF4 but decreased miR-214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR-214 to facilitate ATF4 expression, while miR-214 inhibition and ATF4 overexpression reversed the MEG3 knockdown-mediated decrease in the expression of FoxO1 and FoxO1-downstream targets phosphoenolpyruvate carboxykinase and glucose-6-phosphatase catalytic subunit. In HFD-fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while down-regulating HFD-induced ATF4 expression and upregulating HFD-suppressed miR-214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR-214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 diabetes mellitus.
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