吉林大学学报(医学版)

• 基础研究 • 上一篇    下一篇

电离辐射对破骨细胞分化过程中RANK表达的影响及其致骨损伤的分子机制

周慧2, 杨冰2.3, 唐泉2, 孙元明2, 韩英2, 樊飞跃2,刘晓冬4, 贾立立4   

  1. (1.天津市临床药物关键技术重点实验室,天津医科大学药学院,天津 300070;2. 北京协和医学院&中国医学科学院 放射医学研究所放射生物研究室,天津 300192;3. 天津医科大学基础医学院细胞生物学系,天津 300070;4.吉林大学公共卫生学院 卫生部放射生物学重点实验室,吉林 长春130021)
  • 收稿日期:2013-06-11 发布日期:2013-11-28
  • 通讯作者: 贾立立 E-mail:(Tel:0431-85619443,E-mail:jiall@jlu.edu.cn)
  • 作者简介:周 慧(1983-),女,天津市人,实验师,理学硕士,主要从事分子生物学与药物化学方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(30970867);吉林大学基本科研业务费资助课题(201103082)

Effect of ionizing radiation on expression of RANK in differentiation process of osteoclasts  and its molecular mechanism of bone injury

ZHOU Hui1,YANG Bing2,3,TANG Quan2,SUN Yuan-ming2,HAN Yin2,FAN Fei-yue2,LIU Xiao-dong4,JIA Li-li   

  1. (1. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics andDiagnostics(Theranostics),School of Pharmacy,Tianjin Medical University,Tianjin
     300070,China;2. Radiation Biology Laboratory,Institute of Radiation Medicine Chinese Academy
     of Medical Sciences & Peking Union Medical College,Tianjin 300192,China;3. Department of Cell Biology,College of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China; 4.Key Laboratory of Radiobiology,Ministry of Health,School of Public Health,Jilin University,Chan
    gchun 130021,China)
  • Received:2013-06-11 Published:2013-11-28

摘要:

目的:研究电离辐射对破骨细胞分化过程中核因子κB受体活化因子(RANK) mRNA和蛋白表达的影响,探讨辐射导致骨损伤的分子机制。方法:采用50 μg•L-1 核因子κB受体活化因子配体(RANKL)诱导RAW264.7细胞分化为破骨细胞。RQW264.7细胞分为对照组(未处理)、RANKL处理组(50 μg•L-1RANKL处理)、照射处理组(2 Gy γ射线照射)、照射联合RANKL处理组(50 μg•L-1RANK处理+2 Gy γ射线照射)。采用抗酒石酸磷酸酶(TRAP)染色法检测各组破骨细胞的分化状态;采用PCR法检测各组细胞中RANK mRNA的表达水平;采用Western blotting法检测各组细胞中RANK蛋白的表达水平。 结果:RAW264.7细胞经过RANKL诱导7 d后,TRAP染色呈现阳性,表明已经成功分化成为破骨细胞。与对照组比较,RANKL处理组和照射处理组破骨细胞前体细胞中RANK mRNA和蛋白的表达水平上调(P<0.05);与RANKL处理组比较,照射联合RANKL组破骨细胞前体细胞中RANK mRNA和蛋白表达水平降低(P<0.05)。结论:电离辐射可以促进破骨细胞前体细胞的增殖和成熟,增加其活性,但是对破骨细胞的增殖、成熟与活性却有一定的抑制作用。

关键词: 电离辐射, RAW264.7细胞, 破骨细胞, RANK, 放射治疗

Abstract:

To investigate the effect of ionizing radiation on the expressions of receptor activator for NF-κB(RANK) mRNA and protein in  differentiation process of osteoclasts and to discuss the molecular mechanism of bone injury induced by ionizing radiation.Methods The osteoblasts were differentiated from RAW264.7 cells after treated by 50 μg•L-1 receptor activatorfor NF-κB ligand(RANKL).The RAW264.7 cells were divided into   control group (normal RAW264.7 cells without treatment),RANKL treatment  group(RAW264.7 cells treated with   50 μg•L-1  RANKL), radiation treatment group (RAW264.7 cells exposed to 2 Gy γ-rays) and  radiation combined with RANKL treatment group(RAW264.7 cells treated with both 2 Gy  γ-rays and 50 μg•L-1  RANKL).Tartrate resistant acid phosphatase(TRAP) staining method was used to assess the osteoclast differentiation status;the expression levels of RANK mRNA and protein of  ostoclasts were detected by  PCR method and Western blotting method,respectivly.Results After treated with RANKL for 7 d,the RAW264.7 cells showed TRAP positive result,which indicated the formation of osteoclasts.Compared with control group,the expression levels of  RANK mRNA and protein  of  osteoclast precursors in RANKL treatment group and radiation treatment group  were increased (P<0.05).Compared with RANKL treatment group,the expression levels of RANK mRNA  and protein in radiation  combined  with RANKL  treatment group were decreased(P<0.05). Conclusion Ionizing radiation  may promote the proliferation and mature,and increase the activity of osteoclast precursors,but it  can inhibit the proliferation,mature,and activity of osteoclasts.

Key words: ionizing radiation, RAW264.7, osteoclasts, radiotherapy

中图分类号: 

  • R144.1