Yin Jiaojiao,Zhou Lijun,Wang Yu,Liu Xiaodan,Gu Yongqing,Zhou Pingkun.Effect of salt-inducible kinase 2 on checkpoint in response to γ-ray irradiation[J].Chinese Journal of Radiological Medicine and Protection,2014,34(5):338-341
Effect of salt-inducible kinase 2 on checkpoint in response to γ-ray irradiation
Received:September 23, 2013  
DOI:10.3760/cma.j.issn.0254-5098.2014.05.005
KeyWords:salt-inducible kinase 2  Cell cycle  Histon protein H3  G2/M checkpoint  γ-rays
FundProject:国家自然科学基金(81172119,31370843,31270894)
Author NameAffiliationE-mail
Yin Jiaojiao Anhui Medical University, Hefei 230032, China  
Zhou Lijun 中国人民解放军海军总医院中心实验科  
Wang Yu 军事医学科学院放射与辐射医学研究所  
Liu Xiaodan 军事医学科学院放射与辐射医学研究所  
Gu Yongqing 军事医学科学院放射与辐射医学研究所  
Zhou Pingkun 军事医学科学院放射与辐射医学研究所 zhoupk@bmi.ac.cn 
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Abstract::
      Objective To investigate the effect of salt-induced kinase 2 (SIK2) in the G2/M checkpoint in response to ionizing radiation and the possible mechanism.Methods HeLa cells were irradiated with 60Co γ-rays.The cell model of knockdown SIK2 expression was constrcuted by transfecting HeLa cells with a pSicoR-based lentivirus vector of expressing SIK2 shRNA by lipofectamin 2000.Western blot and flow cytometry were performed to measure the changes of SIK2 protein level and cell cycle distribution.The phosphorylated histone protein H3 on Ser 10 was used as a molecular marker of mitotic cells for detecting the function of G2/M checkpoint.Results The expression level of SIK2 protein increased in HeLa cells after 60Co γ-ray irradiation.A cell model of knockdown SIK2 expression was successfully generated by transfecting the specific shRNA against SIK2.Depression of SIK2 significantly increased the cellular sensitivity at 1, 2, 4, 6 Gy post-irradiation(t=-3.445,-2.581,-3.251,-2.553,P<0.05), and led cells to release earlier from the G2/M boundary arrest compared to control cells at 5, 6 h post-irradiation(t=4.341,6.500,P<0.05).Western blot analysis indicated that the irradiation-induced phosphorylated CHK2/T68 in SIK2 knock-down cells was earlier than that in control cells.Conclusions salt-induced kinase 2 (SIK2) participates in the regulation of G2/M checkpoint induced by ionizing radiation and affects cellular radiosensitivity.
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