RAB32对慢性粒细胞白血病K562细胞增殖和迁移作用的研究

doi:10.12092/j.issn.1009-2501.2019.12.001

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (12): 1321-1327.doi: 10.12092/j.issn.1009-2501.2019.12.001

• 基础研究 • 下一篇

RAB32对慢性粒细胞白血病K562细胞增殖和迁移作用的研究

鲍静^1,2，李小枫¹，章涵硕²，许庆庆¹，孟晓明¹，黄成¹，李俊¹

¹安徽医科大学药学院，合肥 230032，安徽；²安徽医科大学第一附属医院血液科，合肥 230032，安徽

收稿日期:2019-08-09 修回日期:2019-11-12 出版日期:2019-12-26 发布日期:2020-01-07
通讯作者: 李俊，男，博士，教授，博士生导师，研究方向：药理学。 Tel：0551-65161001 E-mail:lj@ahmu.edu.cn
作者简介:鲍静，女，博士，研究方向：临床药理学。 Tel：13956975137 E-mail:doctorbaojing@163.com
基金资助:
安徽高校自然科学研究项目（KJ2019A0233）；国家自然科学基金项目（81770609）

Role of RAB32 on proliferation and migration of chronic myeloid leukemia K562 cells

BAO Jing^1,2，LI Xiaofeng ¹，ZHANG Hanshuo²，XU Qingqing ¹，MENG Xiaoming¹，HUANG Cheng¹，LI Jun¹

¹ Department of Pharmacy, Anhui Medical University, Hefei 230032, Anhui, China; ²Department of Hematology, the First Affiliated Hospital of Anhui Medical University, Hefei 230032, Anhui, China

Received:2019-08-09 Revised:2019-11-12 Online:2019-12-26 Published:2020-01-07

摘要/Abstract

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。

关键词: RAB32, shRNA, 人白血病K562细胞, 增殖, 迁移

Abstract:

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.

Key words: RAB32, shRNA, human leukemia K562 cells, proliferation, migration

中图分类号:

R965.2

鲍静，李小枫，章涵硕，许庆庆，孟晓明，黄成，李俊. RAB32对慢性粒细胞白血病K562细胞增殖和迁移作用的研究[J]. 中国临床药理学与治疗学, 2019, 24(12): 1321-1327.

BAO Jing，LI Xiaofeng，ZHANG Hanshuo，XU Qingqing，MENG Xiaoming，HUANG Cheng，LI Jun. Role of RAB32 on proliferation and migration of chronic myeloid leukemia K562 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(12): 1321-1327.

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RAB32对慢性粒细胞白血病K562细胞增殖和迁移作用的研究

Role of RAB32 on proliferation and migration of chronic myeloid leukemia K562 cells

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