Effects and mechanism of NCTD on proliferation and apoptosis of human leukemic cells by targeting PPP5C
10.3969/j.issn.1671-7856.2024.04.002
- VernacularTitle:NCTD通过调控PPP5C影响人白血病细胞增殖和凋亡的机制研究
- Author:
Xin ZHANG
1
,
2
;
Bingjie CUI
;
Guoxing YU
;
Fei WANG
;
Liang ZHAO
;
Na GAO
;
Jing DU
Author Information
1. 滨州医学院第一临床医学院血液内科,山东滨州 256603
2. 滨州医学院附属医院医学研究中心,山东滨州 256603
- Keywords:
PPP5C;
norcantharidin;
human leukemic cells;
proliferation;
apoptosis
- From:
Chinese Journal of Comparative Medicine
2024;34(4):11-19
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effects and mechanism of norcantharidin(NCTD)on proliferation and apoptosis of NB4 and K562 human leukemic cells by regulating phosphoprotein phosphatase 5 catalytic(PPP5C).Methods PC3.1 and PPP5C-PC3.1 plasmids were electroporated into NB4 and K562 cells.Stable NB4 and K562 cell lines were selected with geneticin(G418).Protein and mRNA expression levels of PPP5C were measured by Western blot and RT-qPCR,respectively.Proliferation,migration,and apoptosis of NB4 and K562 cells were determined by a CCK-8 assay,transwell assay,and Live & Dead? animal cell viability/toxicity detection kit,respectively.NB4 and K562 cells were divided into 0 μg/mL NCTD group and various NCTD dose groups,and cultured in RPMI 1640 medium containing 0,8,16,or 32 μg/ml NCTD.The Live & Dead? animal cell viability/toxicity detection kit measured the numbers of dead and live cells,and cell morphology was observed under a microscope.Western blot was used to measure protein expression levels of caspase 3,Cleaved caspase 3,JNK,p-JNK,p38,p-p38,and α-Tubulin.Results Proliferation,migration,and apoptosis of NB4 and K562 cells were enhanced by overexpression of PPP5C.Compared with 0 μg/mL NCTD group,NCTD promoted apoptosis in a dose-dependent manner.PPP5C overexpression antagonized the killing effect of NCTD on leukemic cells.Mechanistic investigations showed that PPP5C reduced the protein level of p-JNK by dephosphorylating and regulating the expression of apoptosis-related protein Cleaved caspase 3.Conclusions NCTD promotes apoptosis of NB4 and K562 cells and inhibits their proliferation by inhibiting PPP5C.
