GSK3 inhibitors CHIR99021 and 6-bromoindirubin-3'-oxime inhibit microRNA maturation in mouse embryonic stem cells.
Wnt/β-catenin signalling plays a prominent role in maintaining self-renewal and pluripotency of mouse embryonic stem cells (mESCs). microRNAs (miRNAs) have critical roles in maintaining pluripotency and directing reprogramming. To investigate the effect of GSK3 inhibitors on miRNA expression, we analysed the miRNA expression profile of J1 mESCs in the absence or presence of CHIR99021 (CHIR) or 6-bromoindirubin-3'-oxime (BIO) by small RNA deep-sequencing. The results demonstrate that CHIR and BIO decrease mature miRNAs of most miRNA species, 90.4% and 98.1% of the differentially expressed miRNAs in BIO and CHIR treated cells were downregulated respectively. CHIR and BIO treatment leads to a slight upregulation of the primary transcripts of the miR-302-367 cluster and miR-181 family of miRNAs, these miRNAs are activated by Wnt/β-catenin signalling. However, the precursor and mature form of the miR-302-367 cluster and miR-181 family of miRNAs are downregulated by CHIR, suggesting CHIR inhibits maturation of primary miRNA. Western blot analysis shows that BIO and CHIR treatment leads to a reduction of the RNase III enzyme Drosha in the nucleus. These data suggest that BIO and CHIR inhibit miRNA maturation by disturbing nuclear localisation of Drosha. Results also show that BIO and CHIR induce miR-211 expression in J1 mESCs.
1] College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China [2] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Yongyan
Y
Wu
1] College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China [2] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Fayang
F
Liu
1] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China [2] College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China.
Yingying
Y
Liu
1] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China [2] College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China.
Xiaolei
X
Liu
1] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China [2] College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China.
Zhiying
Z
Ai
1] College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China [2] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Zekun
Z
Guo
1] College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China [2] Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Yong
Y
Zhang
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D000066673:Cell Self Renewal / Q000187:drug effects / Q000235:genetics; D048430:Cell Shape / Q000187:drug effects; D003114:Colony-Forming Units Assay; D044127:Epigenesis, Genetic / Q000187:drug effects; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D007211:Indoles / Q000494:pharmacology*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005810:Multigene Family; D010091:Oximes / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D012323:RNA Processing, Post-Transcriptional / Q000187:drug effects* / Q000235:genetics; D043244:Ribonuclease III / Q000378:metabolism; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects / Q000235:genetics; D051176:beta Catenin / Q000378:metabolism
C483321:6-bromoindirubin-3'-oxime; C473711:Chir 99021; D007211:Indoles; D035683:MicroRNAs; D010091:Oximes; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011743:Pyrimidines; D014157:Transcription Factors; D051176:beta Catenin; D038362:Glycogen Synthase Kinase 3; C513400:Drosha protein, mouse; D043244:Ribonuclease III