中国比较医学杂志2024，Vol.34Issue(4)：11-19,9.DOI:10.3969/j.issn.1671-7856.2024.04.002
NCTD通过调控PPP5C影响人白血病细胞增殖和凋亡的机制研究
Effects and mechanism of NCTD on proliferation and apoptosis of human leukemic cells by targeting PPP5C
摘要
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.关键词
PPP5C/去甲斑蝥素/人白血病细胞/增殖/凋亡Key words
PPP5C/norcantharidin/human leukemic cells/proliferation/apoptosis分类
医药卫生引用本文复制引用
张鑫,崔冰洁,于国兴,王飞,赵靓,高娜,杜静..NCTD通过调控PPP5C影响人白血病细胞增殖和凋亡的机制研究[J].中国比较医学杂志,2024,34(4):11-19,9.基金项目
国家自然科学基金(82373097,31900441) （82373097,31900441）
山东省自然科学基金(ZR2019MC026) （ZR2019MC026）
山东省中医药发展计划(2019-0514) （2019-0514）
泰山学者青年专家经费(tsqn202103191) （tsqn202103191）
滨州医学院科研计划与科研启动基金项目(BY2022KJ64). （BY2022KJ64）
