中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (27): 5062-5066.doi: 10.3969/j.issn.2095-4344.2012.27.023

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

姜黄素逆转SP600125诱导Dami细胞多倍体化模型的建立

李艳平1，周 凡2，马东初3，姜志明3，刘彦琴2，刘景华2，王吉刚2   

  1. 1辽宁医学院中国人民解放军沈阳军区总医院研究生培养基地，辽宁省锦州市 121001；解放军沈阳军区总医院， 2血液科，3医学实验科，辽宁省沈阳市 110016
  • 收稿日期:2012-01-11 修回日期:2012-03-25 出版日期:2012-07-01 发布日期:2013-11-01
  • 通讯作者: 周凡，教授，硕士研究生导师，解放军沈阳军区总医院血液科，辽宁省沈阳市 110016 zhoufan611@sina.com
  • 作者简介:李艳平★，女，1983年生，河北省唐山市人，汉族，辽宁医学院在读硕士，医师，主要从事血液肿瘤及免疫治疗方面的研究。 liyanping0627@163.com

Establishment of Dami cell model of SP600125-induced polyploidization which was reversed by curcumin

Li Yan-ping1, Zhou Fan2, Ma Dong-chu3, Jiang Zhi-ming3, Liu Yan-qin2, Liu Jing-hua2, Wang Ji-gang2   

  1. 1Postgraduate Culture Base of the General Hospital of Shenyang Military Region, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China;
    2 Department of Hematology, 3Department of Laboratory Medicine, the General Hospital of Shenyang Military Region, Shenyang 110016, Liaoning Province, China
  • Received:2012-01-11 Revised:2012-03-25 Online:2012-07-01 Published:2013-11-01
  • Contact: Zhou Fan, Professor, Master’s supervisor, Department of Hematology, the General Hospital of Shenyang Military Region, Shenyang 110016, Liaoning Province, China zhoufan611@sina.com

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摘要：

背景：细胞的多倍性是重要的生物学现象，不仅影响组织器官和造血系统的正常生理功能，而且是某些应激条件下代偿性的表现，和发生某些病理过程的基础，尤其与恶性肿瘤的发生有着密切的关系。
目的：优化SP600125诱导Dami细胞多倍体化模型，探讨姜黄素对此多倍体化逆转作用的机制。
方法：将不同浓度SP600125(15，30，60 mmol/L)分别加入细胞悬液中，诱导Dami细胞多倍体化，确定最佳诱导时间和浓度；将5，10，20 mmol/L姜黄素分别加入含有SP600125的细胞悬液中，培养72 h收集细胞。Annexin V-PI染色法的流式细胞术检测细胞倍性的变化、Western blot法检测细胞周期相关蛋白的变化。
结果与结论：30 mmol/L SP600125作用Dami细胞72 h，诱导的多倍体化细胞数量最多，并且cyclin D3表达最高；姜黄素明显逆转Dami细胞多倍体化，cyclin D3表达逐渐降低，存在明显的剂量依赖关系。结果显示，实验成功建立了Dami细胞的多倍体化模型，并确立了诱导多倍体化Dami细胞的SP600125最佳诱导时间和浓度72 h，30 mmol/L。姜黄素可能通过抑制cyclin D3的表达，逆转SP600125诱导Dami细胞多倍体化。

关键词: 姜黄素, SP600125, 巨核细胞, 多倍体化, 细胞周期蛋白, 干细胞

Abstract:

BACKGROUND: Cell polyploidization is an important biological phenomenon. It not only influences the normal physiological function of tissue and hematopoietic system, but also is the compensatory performance under stress condition. It is closely related to the occurrence of some pathological processes, in particular malignant tumorigenesis.
OBJECTIVE: To optimize the Dami cell model of SP600125-induced polyploidization and to investigate the mechanism by which curcumin reversed the polyploidization.
METHODS: 15, 60, 60 μmol/mL SP600125 was added to the cell suspension to induce the polyploidization of Dami cells. The optimal induction time and concentration were determined. Then 5, 10, 20 mmol/L curcumin was added to the cell suspension containing SP600125. Cells were collected after 72 hours of culture. Cell ploidy changes were determined by flow cytometry using Annexin V-PI staining. Cell cycle-related protein expression changes were detected by Western blot method.
RESUITS AND CONCLUSION: After treated with 30 mmol/L SP600125 for 72 hours, the number of polyploid Dami cells was increased and the expression of cyclin D3 was highest. The ployploidization of Dami cells was significantly reversed by curcumin, and the expression of cyclin D3 was gradually decreased, showing a significant time- and dose-dependent manner. A Dami cell model of polyploidization was successfully established and the optical induction time and concentration for SP600125 were determined at 72 hours and 30 mmol/L. The polyploidization of Dami cells may be reversed through inhibiting cyclinD3 expression.

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