Aurora kinase-B silencing promotes apoptosis of osteosarcoma 143B cells by ULK1 phosphorylation-induced autophagy.
10.12122/j.issn.1673-4254.2020.09.08
- Author:
Xin WU
1
;
Jiaming LIU
1
;
Honghai SONG
1
;
Qikun YANG
1
;
Hui YING
1
;
Zhili LIU
1
Author Information
1. Spine and Spinal Cord Disease Centre, First Affiliated Hospital of Nanchang University, Institute of Spine and Spinal Cord Disease, Nanchang University, Nanchang 330006, China.
- Publication Type:Journal Article
- Keywords:
Aurora kinase B;
ULK1 protein;
apoptosis;
autophagy;
osteosarcoma
- From:
Journal of Southern Medical University
2020;40(9):1273-1279
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of Aurora kinase B (AURKB) silencing-induced autophagy on apoptosis of osteosarcoma 143B cells and the underlying molecular mechanisms.
METHODS:Human osteosarcoma 143B cells were transfected with Lv/shAURKB or the negative control vector Lv/shScrambled followed by treatment with chloroquine (CQ) for 24 h. Western blotting was performed to detect the protein expression levels of AURKB, P62, LC3, cleaved caspase-3, Bcl-2, and P-ULK1. Transmission electron microscopy and LC3 dual-label fluorescence method were used to trace the autophagosomes in 143B cells to assess cell autophagy, and the cell apoptosis was detected using flow cytometry and TUNEL assay. Co-immunoprecipitation assay was used to detect the interaction between AURKB and ULK1.
RESULTS:The ratio of autophagy-related proteins LC3 II/I and the number of autophagosomes were significantly increased in 143B cells after transfection with Lv/shAURKB ( < 0.05), which significantly increased the expression of cleaved caspase-3 and reduced the expression of Bcl-2 ( < 0.05). Combined treatment of the cells with Lv/shAURKB and the autophagy inhibitor chloroquine obviously restored the expressions of caspase-3 and Bcl-2 ( < 0.05). Transfection with Lv/shAURKB significantly increased the apoptosis rate of 143B cells ( < 0.05), and this effect was significantly antagonized by combined treatment with chloroquine ( < 0.05). AURKB silencing strongly activated the phosphorylation of the autophagy-initiating protein ULK1 in 143B cells ( < 0.05). The results of co-immunoprecipitation assay confirmed when AURKB was immunoprecipitated, ULK1 also precipitated.
CONCLUSIONS:Silencing AURKB can induce autophagy by activating ULK1 phosphorylation to promote apoptosis in 143B cells.
