Knockdown of AMPK?2 impairs epithelial?mesenchymal transition in rat renal tubular epithelial cells by downregulating ETS1 and RPS6KA1.

ABSTRACT: Epithelial?mesenchymal transition (EMT) serves an important regulatory role in obstructive nephropathy and renal fibrosis. As an intracellular energy sensor, AMP?activated protein kinase (AMPK) is essential in the process of EMT. The aim of the present study was to elucidate changes in the expression levels of AMPK?2 and which AMPK?2 genes play a role during EMT. TGF??1 was used to induce EMT in normal rat renal tubular epithelial (NRK?52E) cells. The short hairpin AMPK?2 lentivirus was used to interfere with AMPK?2 expression levels in EMT?derived NRK?52E cells and AMPK?2 expression levels and EMT were detected. Differential gene expression levels following AMPK?2 knockdown in EMT?derived NRK?52E cells were assessed via gene microarray. Potential regulatory pathways were analyzed using ingenuity pathway analysis (IPA) and differentially expressed genes were partially verified by reverse transcription?quantitative PCR (RT?qPCR) and western blotting. AMPK?2 was upregulated in TGF??1?induced EMT?derived NRK?52E cells. EMT progression was significantly inhibited following downregulation of expression levels of AMPK?2 by shAMPK?2 lentivirus. A total of 1,588 differentially expressed genes were detected following AMPK?2 knockdown in NRK?52E cells in which EMT occurred. The ERK/MAPK pathway was significantly impaired following AMPK?2 knockdown, as indicated by IPA analysis. Furthermore, RT?qPCR and western blot results demonstrated that the expression levels of AMPK?2, v?ets erythroblastosis virus E26 oncogene homolog?1 (ETS1) and ribosomal protein S6 kinase A1 (RPS6KA1) were upregulated following EMT in NRK?52E cells, whereas the expression levels of ETS1 and RPS6KA1 were downregulated following AMPK?2 knockdown. It was concluded that AMPK?2 plays a key role in the regulation of rat renal tubular EMT, which may be achieved by modulating ETS1 and RPS6KA1 in the ERK/MAPK pathway.

SUBMITTER: Yin X 

PROVIDER: S-EPMC7646838 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature