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 Accumulation of Intracellular Drug of HL60/A and HL60 Cell and Its Mechanism

作　　者：陈小芳[1] 陈显凌[2] 陈元仲[2] CHEN Xiao-Fang;CHEN Xian-Ling;CHEN Yuan-Zhong(Department of Infectious Disease,Fujian Medical University Union Hospital;Fujian Inastitute of Hematology,Fujian Provincial Key Laboratory on Hematology,Fujian Medical University Union Hospital,Fuzhou 350001,Fujian Province,China)

机构地区：[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

出　　处：《中国实验血液学杂志》2024年第1期71-77,共7页Journal of Experimental Hematology

基　　金：福建省科技创新联合基金(2017Y9054);福建省自然科学基金(2022J01257)。

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。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 

分 类 号：R733.71[医药卫生—肿瘤]