摘要
目的:探讨RAB32对慢性粒细胞白血病(CML)K562细胞增殖和迁移作用的影响。方法:使用Western blot和qRT-PCR分别检测RAB32在CML患者、健康对照以及CML K562细胞株中的表达情况。进一步转染RAB32-shRNA到K562细胞株中敲除RAB32基因后,流式细胞仪分析K562细胞周期变化,Western blot和qRT-PCR分别检测K562细胞中细胞增殖相关蛋白C-myc、Cyclin D1和细胞迁移相关蛋白MMP-3、MMP-9的蛋白和mRNA表达水平变化。结果:Western blot和qRT-PCR检测结果显示,RAB32在CML患者和K562细胞株中高表达(P<0.01)。在K562细胞株中转染RAB32-shRNA抑制RAB32表达后,细胞周期分析结果显示,较NC-shRNA组相比,RAB32-shRNA组K562细胞的S期和G2/M期细胞比例均有不同程度降低,且S期细胞比例降低更为显著(P<0.01)。Western blot检测结果显示,与NC-shRNA组相比,RAB32-shRNA组K562细胞的细胞增殖相关蛋白C-myc、Cyclin D1和细胞迁移相关蛋白MMP-3、MMP-9蛋白表达水平明显降低(P<0.01),qRT-PCR检测结果显示,这四种因子的mRNA水平也相应降低(P<0.01)。结论:抑制RAB32的表达能够抑制CML K562细胞的增殖和迁移能力。
AIM:To investigate the effects of RAB32 on the proliferation and migration of chronic myeloid leukemia(CML)K562 cells.METHODS:The expression of RAB32 in newly diagnosed CML patients,healthy volunteers and K562 cell lines were assessed using Western blot and qRT-PCR.In addition,RAB32 gene expression was knocked down using RAB32-shRNA,and then the cell cycle variation of K562 cell was analyzed by flow cytometry.The expressions of cell proliferation-related genes(C-myc and CyclinD1)and migration-associated genes(MMP-3 and MMP-9)were assessed using Western blot and qRT-PCR.RESULTS:The results of Western blot and qRT-PCR showed that RAB32 were highly expressed in both CML patients and the K562 cell line(P<0.01).Cell cycle analysis showed that compared with NC-shRNA group,the RAB32-shRNA group had decreased proportions of S-phase and G2/M-phase,especially S-phase cells(P<0.01).The protein levels of C-myc,Cyclin D1,MMP-3 and MMP-9 were significantly down-regulated in K562 cells transfected with RAB32-shRNA compared with NC-shRNA group(P<0.01).The mRNA levels of these four factors were also reduced in RAB32-shRNA group compared to NC-shRNA group(P<0.01).CONCLUSION:Down-regulation of RAB32 could weaken K562 cell proliferation and migration.
作者
鲍静
李小枫
章涵硕
许庆庆
孟晓明
黄成
李俊
BAO Jing;LI Xiaofeng;ZHANG Hanshuo;XU Qingqing;MENG Xiaoming;HUANG Cheng;LI Jun(Department of Pharmacy, Anhui Medical University, Hefei 230032, Anhui, China;Department of Hematology, the First Affiliated Hospital of Anhui Medical University, Hefei 230032, Anhui, China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2019年第12期1321-1327,共7页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
安徽高校自然科学研究项目(KJ2019A0233)
国家自然科学基金项目(81770609)
