Preliminary Study on the Effect of Silencing Nucleostemin Com- bined with Rapamycin on Autophagy and Apoptosis of HL-60 Cells.
10.19746/j.cnki.issn.1009-2137.2023.06.005
- Author:
Ya-Qi WANG
1
,
2
;
Xiao-Juan GAO
1
,
2
;
Bao-Hong YUE
3
,
4
Author Information
1. Department of Laboratory Medicine , The First Afliated Hospital of Zhengzhou University , Key Clinical Laboratory of Henan Province , Faculty of Laboratory Medicine , Zhengzhou University
2. Zhengzhou 450052 , Henan Prorince , China.
3. Department of Laboratory Medicine , The First Afliated Hospital of Zhengzhou University , Key Clinical Laboratory of Henan Province , Faculty of Laboratory Medicine ,Zhengzhou University
4. Zhengzhou 450052 , Henan Prorince , China E-mail: ybh2002@ 163 . com.
- Publication Type:Journal Article
- Keywords:
HL-60 cells;
nucleostemin;
apoptosis;
autophagy;
rapamycin
- MeSH:
Humans;
HL-60 Cells;
Sirolimus/pharmacology*;
bcl-2-Associated X Protein;
Autophagy;
Apoptosis
- From:
Journal of Experimental Hematology
2023;31(6):1629-1634
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects of knocking down nucleostemin ( NS) combined with rapamycin (RAPA) on autophagy and apoptosis in HL-60 cells , and to explore its role in HL-60 cells .
METHODS:The expression of NS protein was detected using Western blot , after transfection of HL-60 cells was achieved by the recombinant lentviral vector NS -RNAi-GV248 . Flow cytometry was used to detect changes in cells apoptosis after NS silencing/ rapamycin for 24 , 48 hours , and the expressions of NS , LC3 , p62 , BCL-2 and Bax proteins in cells were detected by Western blot.
RESULTS:The expression of NS in HL-60 cells was successfully down-regulated by recombinant lentiviral vector. After treatment with rapamycin for 24 and 48 h , the apoptosis rate of cells in each group increased (P < 0.05) , and the apoptosis was more obvious at 48 hours . Compared with the NS silencing group or rapamycin group , after treated with NS down-regulation combined with rapamycin for 48 hours , the apoptosis of HL-60 cells was significantly increased ( P < 0.05 ) , LC3 -II/LC3 -I ratio was significantly increased ( P < 0.05 ) , p62 protein expression was significantly decreased (P < 0.05) , and BCL-2/Bax ratio was significantly decreased ( P < 0.05) .
CONCLUSION:NS down-regulation combined with rapamycin can enhance the apoptosis and autophagy of HL-60 cells , and the induction of apoptosis of HL-60 cells may be related to the expression of BCL-2 and Bax proteins .
