TCP1表达对HL60和HL60/A细胞增殖及细胞内药物蓄积的调控作用及其机制
The Effect of TCP1 Expression on the Proliferation and the Accu-mulation of Intracellular Drug of HL60/A and HL60 Cell and Its Mechanism
陈小芳 1陈显凌 2陈元仲2
作者信息
- 1. 福建医科大学附属协和医院感染科
- 2. 福建医科大学附属协和医院血液科,福建省血液病学重点实验室,福建省血液病研究所,福建福州 350001
- 折叠
摘要
目的:研究TCP1表达对HL60及HL60/A细胞增殖及细胞内药物蓄积的影响及其机制.方法:利用慢病毒转染技术构建敲低、过表达TCP1的HL60/A细胞和HL60细胞及其对照组细胞,Western blot评估敲低、过表达效率.采用CCK-8法检测细胞增殖能力;激光共聚焦显微镜及流式细胞术检测细胞内的药物蓄积;流式细胞术及Western blot检测膜转运蛋白(MRP1、P-gP)及p-AKT的表达水平.结果:在HL60/A细胞中敲低TCP1的表达能够抑制细胞增殖,增加细胞内药物蓄积,降低转运蛋白MRP1及P-gP的表达,在HL60细胞中过表达TCP1能够促进细胞的增殖,减少细胞内药物蓄积,提高转运蛋白MRPI及P-gP的表达.同时利用PI3K抑制剂LY294002抑制PI3K/AKT信号能够拮抗TCP1过表达所致的细胞增殖活性增强、细胞内药物蓄积减少及MRP1、P-gP表达的升高.结论:TCP1能够促进细胞增殖,并通过激活PI3K/AKT信号促进转运蛋白MRP1、P-gP的表达,降低细胞内的药物蓄积.
Abstract
Objective:To investigate the effect of TCP1 expression on the proliferation and the accumulation of intracellular drug of HL60/A and HL60 cells and its possible molecular mechanism.Methods:Lentiviral transfection technology was used to construct HL60/A and HL60 cells with knocked down or overexpressed TCP1 and their control cells.The efficiency of knockdown and overexpression was evaluated by Western blot.The cell proliferation was detected by CCK-8 assay.The intracellular drug accumulation was detected by laser confocal detection and flow cytometry.The expression levels of MRP1,P-gP and p-AKT were evaluated by flow cytometry and Western blot.Results:After TCP1 was knocked down,the proliferation ability of HL60/A cells was significantly reduced,the accumulation of intracellular drug was significantly increased and the expression of MRP1 and P-gP protein were decreased.After TCP1 was overexpressed,the proliferation ability of HL60 was significantly increased,the accumulation of intracellular drug was significantly decreased and the expression of MRP1 and P-gP protein were increased.Intervention of LY294002 significantly antagonized the promotion on cell proliferation,the inhibition on intracellular drug accumulation and the expression of MRP1 and P-gP mediated by TCP1 overexpressing in HL60 cells.Conclusion:TCP1 can promote cell proliferation,improve the expression of MRP1 and P-gP by activating PI3K/AKT signal,and reduce intracellular drug accumulation.
关键词
TCP1/急性髓系白血病/药物蓄积/MRP1/P-gP/PI3K/AKT信号通路Key words
TCP1/acute myeloid leukemia/intracellular drug accumulation/MRP1/P-gP/PI3K/AKT signaling pathway引用本文复制引用
基金项目
福建省科技创新联合基金(2017Y9054)
福建省自然科学基金(2022J01257)
出版年
2024
NETL
NSTL
万方数据