Effect of Down-Regulation of ANRIL on Proliferation and Apoptosis of Kasumi-1 Cells and Its Potential Mechanism.
10.19746/j.cnki.issn.1009-2137.2022.04.002
- Author:
Cheng-Si ZHANG
1
;
Jian-Xia XU
2
;
Fa-Hua DENG
2
;
Hua-Li HU
2
;
Si-Qi WANG
2
;
Hai HUANG
3
;
Si-Xi WEI
4
Author Information
1. School of Clinical Laboratory Science, Guizhou Medical University, Guizhou Center for Disease Control and Prevention.
2. School of Clinical Laboratory Science, Guizhou Medical University Guiyang 550004, Guizhou Province, China.
3. School of Clinical Laboratory Science, Guizhou Medical University Center for Clinical Laboratories, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China.
4. School of Clinical Laboratory Science, Guizhou Medical University Center for Clinical Laboratories, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China. E-mail: wsixi@gmc.edu.cn.
- Publication Type:Journal Article
- Keywords:
ANRIL;
PI3K/AKT;
apoptosis;
proliferation
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Down-Regulation;
Humans;
Leukemia, Myeloid, Acute/genetics*;
Phosphatidylinositol 3-Kinases/metabolism*;
Proto-Oncogene Proteins c-akt/genetics*;
RNA, Long Noncoding/genetics*
- From:
Journal of Experimental Hematology
2022;30(4):984-989
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the down-regulation of ANRIL (Antisense non-coding RNA in the INK4 Locus) effects on proliferation and apoptosis of Kasumi-1 cells and its related molecular mechanism.
METHODS:Recombinant lentivirus was used to construct ANRIL down-regulation Kasumi-1 cells (sh-ANRIL group) and its control cells (sh-NC group). A fluorescence microscope was used to observe the transfection efficiency, RT-qPCR was used to detect knockdown efficiency and ANRIL expression in PBMCs and MBMCs of patients with acute myeloid leukemia (AML). Proliferation and apoptosis of Kasumi-1 cells were assayed by CCK-8 method and flow cytometry. Western blot was employed to detect the expression of PI3K, AKT, p-AKT, and relevant protein after down-regulation of ANRIL in Kasumi-1 cells.
RESULTS:ANRIL overexpressed significantly in PBMCs and MBMCs of patients with AML, the transfection efficiency of recombinant lentivirus carrying sh-ANRIL and sh-NC on Kasumi-1 cells exceeded 90%, and the knockdown efficiency was 70%. When DNR was administrated for 24, 48, and 72 hours, the cell inhibition rate of the sh-ANRIL group was (47.40±1.49)%, (69.11±0.51)% and (91.82±1.10)%, which were significantly higher than those of the sh-NC group, respectively (P<0.05). The apoptotic rate in the sh-ANRIL group was (10.29±0.58)%, which was significantly higher than (5.42±0.67)% of the sh-NC group (P<0.01). After DNR treatment for 24 hours, the apoptotic rate of the sh-ANRIL group was (54.41±1.69)%, which was significantly higher than (38.28±1.42)% of sh-NC group (P<0.001). Western blot revealed that the protein levels of PI3K, p-AKT, PCNA, and BCL-2 in the sh-ANRIL group were reduced significantly than those in the sh-NC group, while the BAX protein expression increased.
CONCLUSION:ANRIL may affect the proliferation and apoptosis of Kasumi-1 cells through PI3K/AKT signaling pathway. ANRIL is a potential therapeutic target for AML.
