PMID- 33251211
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20240330
IS  - 2296-634X (Print)
IS  - 2296-634X (Electronic)
IS  - 2296-634X (Linking)
VI  - 8
DP  - 2020
TI  - High Expression of miR-204 in Chicken Atrophic Ovaries Promotes Granulosa Cell 
      Apoptosis and Inhibits Autophagy.
PG  - 580072
LID - 10.3389/fcell.2020.580072 [doi]
LID - 580072
AB  - Chicken atrophic ovaries have decreased volume and are indicative of ovarian 
      failure, presence of a tumor, or interrupted ovarian blood supply. Ovarian tumor 
      is accompanied by an increase in follicular atresia, granulosa cell (GC) 
      apoptosis, and autophagy. In a previous study, we found using high throughput 
      sequencing that miR-204 is highly expressed in chicken atrophic ovaries. Thus, in 
      the present study, we further investigated its function in GC apoptosis and 
      autophagy. We found that overexpression of miR-204 reduced mRNA and protein 
      levels of proliferation-related genes and increased apoptosis-related genes. Cell 
      counting kit-8 (CCK-8), 5-ethynyl-2-deoxyuridine (EdU), and flow cytometry assays 
      revealed that miR-204 inhibited GC proliferation and promoted apoptosis. 
      Furthermore, we confirmed with reporter gene assays that Forkhead box K2 (FOXK2) 
      was directly targeted by miR-204. FOXK2, as a downstream regulator of 
      phosphoinositide 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) signal 
      pathways, promoted GC proliferation and inhibited apoptosis. Subsequently, we 
      observed that miR-204 was involved in GC autophagy by targeting Transient 
      Receptor Potential Melastatin 3 (TRPM3). The luciferase activities of the two 
      binding sites of TRPM3 were decreased in response to treatment with a miR-204 
      mimic, and the autophagic flux was increased after miR-204 inhibition. However, 
      overexpression of miR-204 had opposite results in autophagosomes and 
      autolysosomes. miR-204 inhibits GC autophagy by suppressing the protein 
      expression of TRPM3/AMP-activated protein kinase (AMPK)/ULK signaling pathway 
      components. Inhibition of miR-204 enhanced autophagy by accumulating and 
      degrading the protein levels of LC3-II (Microtubule Associated Protein Light 
      Chain 3B) and p62 (Protein of 62 kDa), respectively, whereas miR-204 
      overexpression was associated with contrary results. Immunofluorescence staining 
      showed that there was a significant reduction in the fluorescent intensity of 
      LC3B, whereas p62 protein was increased after TRPM3 silencing. Collectively, our 
      results indicate that miR-204 is highly expressed in chicken atrophic ovaries, 
      which promotes GC apoptosis via repressing FOXK2 through the PI3K/AKT/mTOR 
      pathway and inhibits autophagy by impeding the TRPM3/AMPK/ULK pathway.
CI  - Copyright (c) 2020 Cui, Liu, Kwame Amevor, Zhu, Wang, Li, Shu, Tian and Zhao.
FAU - Cui, Zhifu
AU  - Cui Z
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Liu, Lingbin
AU  - Liu L
AD  - College of Animal Science and Technology, Southwest University, Chongqing, China.
FAU - Kwame Amevor, Felix
AU  - Kwame Amevor F
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Zhu, Qing
AU  - Zhu Q
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Wang, Yan
AU  - Wang Y
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Li, Diyan
AU  - Li D
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Shu, Gang
AU  - Shu G
AD  - Department of Pharmacy, College of Veterinary Medicine, Sichuan Agricultural 
      University, Chengdu, China.
FAU - Tian, Yaofu
AU  - Tian Y
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
FAU - Zhao, Xiaoling
AU  - Zhao X
AD  - Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of 
      Sichuan Province, Sichuan Agricultural University, Chengdu, China.
LA  - eng
PT  - Journal Article
DEP - 20201105
PL  - Switzerland
TA  - Front Cell Dev Biol
JT  - Frontiers in cell and developmental biology
JID - 101630250
PMC - PMC7676916
OTO - NOTNLM
OT  - apoptosis
OT  - atrophic ovary
OT  - autophagy
OT  - chicken
OT  - miR-204
OT  - signaling pathway
EDAT- 2020/12/01 06:00
MHDA- 2020/12/01 06:01
PMCR- 2020/01/01
CRDT- 2020/11/30 05:54
PHST- 2020/07/04 00:00 [received]
PHST- 2020/10/12 00:00 [accepted]
PHST- 2020/11/30 05:54 [entrez]
PHST- 2020/12/01 06:00 [pubmed]
PHST- 2020/12/01 06:01 [medline]
PHST- 2020/01/01 00:00 [pmc-release]
AID - 10.3389/fcell.2020.580072 [doi]
PST - epublish
SO  - Front Cell Dev Biol. 2020 Nov 5;8:580072. doi: 10.3389/fcell.2020.580072. 
      eCollection 2020.