紫杉醇调控NF-κB通路对FLT3-ITD突变急性髓细胞白血病细胞株MOLM-13增殖和凋亡的影响  

Effects of paclitaxel on the proliferation and apoptosis of FLT3-ITD mutant acute myelocytic leukemia cell line MOLM-13 by regulation of NF-κB pathway

作　　者：周宝文 吴梅青 石泽延[1] 白子文 姚奕斌[1] 刘振芳[1] 庞如利 赵卫华[1] Zhou Baowen;Wu Meiqing;Shi Zeyan;Bai Ziwen;Yao Yibin;Liu Zhenfang;Pang Ruli;Zhao Weihua(Department of Hematology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院血液内科,南宁530021

出　　处：《广西医科大学学报》2022年第3期351-356,共6页Journal of Guangxi Medical University

基　　金：国家青年科学基金资助项目(No.81700172);中国博士后科学基金第67批面上项目(No.2020M673097);广西壮族自治区卫生健康委员会自筹经费科研课题(No.Z20200961)。

摘　　要：目的:研究紫杉醇(PTX)对FLT3-ITD突变急性髓细胞白血病细胞株MOLM-13增殖与凋亡的影响及作用机制。方法:不同浓度的PTX分别作用于FLT3-ITD突变细胞MOLM-13和FLT3未突变细胞THP-1,采用CCK8法及流式细胞仪FITC/PI双标法分别检测PTX对两种细胞株增殖与凋亡的影响;采用实时荧光定量PCR法(RT-qPCR)及western blotting法分别检测PTX对 MOLM-13 细胞株 FLT3、NF-κB 基因及蛋白表达的影响。结果:PTX 作用于 MOLM-13 细胞 24 h、48 h、72 h 的 IC_(50)分别为7.659 nmol/L、3.748 nmol/L、3.408 nmol/L,而 THP-1 细胞的 IC_(50)为 61.164 nmol/L、11.804 nmol/L、9.834 nmol/L。4 nmol/L 和8 nmol/L PTX 作用 MOLM-13 细胞 48 h 的凋亡率分别为(13.12±0.20)%、(46.27±1.48)%,与对照组[(9.95±0.48)%]相比凋亡率升高(P<0.05);4 nmol/L和8 nmol/L PTX作用THP-1细胞48 h后的凋亡率分别为(4.26±2.06)%、(8.22±0.47)%,8 nmol/L PTX处理组凋亡率比对照组[(5.26±0.83)%]高(P<0.05)。经 PTX 处理 48 h 后,MOLM-13 细胞中 FLT3、NF-κB 基因表达下调,FLT3、NF-κB、p-NF-κB 蛋白表达下调(P<0.05)。结论:与 FLT3 未突变白血病细胞株 THP-1 相比,PTX 对 FLT3-ITD 突变细胞株MOLM-13的抑制作用更为明显,且呈剂量和时间依赖性;PTX对MOLM-13细胞的作用机制可能与下调FLT3、NF-κB以及p-NF-κB 蛋白的表达,进而诱导白血病细胞发生凋亡有关。Objective:To study the effects of paclitaxel (PTX) on the proliferation and apoptosis of FLT3-ITD mutant acute myeloid leukemia cell line MOLM- 13 and its mechanism.Methods:Different concentrations of PTX were applied to FLT3-ITD mutant cells MOLM-13 and FLT3 mutation-free cells THP-1,and the effects of PTX on the proliferation and apoptosis of the two cell lines were detected by CCK8 assay and flow cytometry FITC/PI double- labeling.The effects of PTX on the mRNA and protein expressions of FLT3 and NF- κB in MOLM-13 cells were detected by RT-qPCR and western blotting.Results:The IC_(50) of PTX acting on MOLM-13cells at 24 h,48 h and 72 h were 7.659 nmol/L,3.748 nmol/L and 3.408 nmol/L respectively,while the IC_(50) of THP-1 cells were 61.164 nmol/L,11.804 nmol/L and 9.834 nmol/L respectively.The apoptosis rates of 4 nmol/L and 8 nmol/L PTX-treated MOLM-13 cells at 48 h were (13.12±0.20)% and (46.27±1.48)% respectively,and the apoptosis rates were increased compared with the control group [(9.95±0.48)%,P<0.05].The apoptosis rates of 4 nmol/L and 8 nmol/L PTX- treated THP-1 cells at 48 h were (4.26±2.06)% and (8.22±0.47)% respectively,and the apoptosis rate was significantly higher in the 8 nmol/L PTX-treated group compared with the control group [(5.26±0.83)%,P<0.05].After 48 h of PTX treatment,the mRNA expressions of FLT3 and NF- κB were down- regulated in MOLM- 13cells (P<0.05).FLT3,NF-κB and p-NF-κB protein expressions were lower in MOLM-13 cells than those in the control group after 48 h of PTX treatment (P<0.05).Conclusion:Compared with the FLT3 mutation-free leukemia cell line THP- 1,the inhibitory effect of PTX on the FLT3- ITD mutant cell line MOLM- 13 is more pronounced and dose- and time-dependent;the mechanism of PTX acting on MOLM-13 cells may be related to the down-regulation of FLT3,NF-κB,and p-NF-κB protein expressions,which in turn is associated with the induction of leukemia cell apoptosis.

关 键 词：紫杉醇 急性髓系白血病 FLT3-ITD NF-ΚB 

分 类 号：R733.71[医药卫生—肿瘤]