Transcriptional Repression of CYP3A4 by Increased miR-200a-3p and miR-150-5p Promotes Steatosis in vitro

Zhijun Huang; Mengyao Wang; Li Liu; Jinfu Peng; Chengxian Guo; Xiaoping Chen; Lu Huang; Jieqiong Tan; Guoping Yang

doi:10.3389/fgene.2019.00484

Transcriptional Repression of CYP3A4 by Increased miR-200a-3p and miR-150-5p Promotes Steatosis in vitro

Front Genet. 2019 May 28:10:484. doi: 10.3389/fgene.2019.00484. eCollection 2019.

Authors

Zhijun Huang^{1

2}, Mengyao Wang^{1

3}, Li Liu¹, Jinfu Peng^{1

2}, Chengxian Guo¹, Xiaoping Chen⁴, Lu Huang^{1

2}, Jieqiong Tan⁵, Guoping Yang^{1

2}

Affiliations

¹ Center for Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, China.
² Department of Pharmacy, The Third Xiangya Hospital, Central South University, Changsha, China.
³ Xiangya School of Pharmaceutical Sciences, Central South University, Changsha, China.
⁴ Institute of Clinical Pharmacology, Central South University, Changsha, China.
⁵ Center for Medical Genetics, Life Science School, Central South University, Changsha, China.

Abstract

Hepatic cytochrome P450 enzyme activities correlate with non-alcoholic fatty liver disease (NAFLD) and hepatic steatosis. The decreased activity of CYP3A4, an important drug-metabolizing enzyme, is associated with the progression of NAFLD. CYP3A4 is predicted as a target gene of miR-200a-3p and miR-150-5p by MicroInspector and TargetScan algorithms analyses. Here, we found decreased CYP3A4 and increased miR-200a-3p and miR-150-5p in LO2 cells with free fatty acid (FFA)-induced steatosis. Dual-luciferase assay confirmed that both miR-200a-3p and miR-150-5p targeted the 3'-untranslated region (3'-UTR) of CYP3A4 and that such interaction was abolished by miRNA binding site mutations in 3'-UTR of CYP3A4. Using miR-200a-3p and miR-150-5p mimics and inhibitors, we further confirmed that endogenous CYP3A4 was regulated posttranscriptionally by miR-200a-3p or miR-150-5p. Moreover, miR-200a-3p and miR-150-5p inhibitors attenuated FFA-induced steatosis in LO2 cells, and such effect was dependent on CYP3Y4 expression. These results suggest that miR-200a-3p and miR-150-5p, through directly targeting 3'-UTR of CYP3A4, contribute to the development of FFA-induced steatosis.

Keywords: CYP3A4; LO2 cell line; miR-150-5p; miR-200a-3p; non-alcoholic fatty liver disease.