[1]李平,宗天舟,季晓芹,等.CD133+U87人脑胶质瘤干细胞放射敏感性和DNA双链断裂损伤修复的实验研究[J].国际放射医学核医学杂志,2013,37(3):153-159.[doi:10.3760/cma.j.issn.1673-4114.2013.03.007]
　LI Ping,ZONG Tian-zhou,JI Xiao-qin,et al.CD133 positive U87 glioma stem cell radiosensitivity and DNA double-strand break repair[J].International Journal of Radiation Medicine and Nuclear Medicine,2013,37(3):153-159.[doi:10.3760/cma.j.issn.1673-4114.2013.03.007]
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CD133+U87人脑胶质瘤干细胞放射敏感性和DNA双链断裂损伤修复的实验研究(/HTML)
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《国际放射医学核医学杂志》[ISSN:1673-4114/CN:12-1381/R]

卷:
37
期数:
2013年第3期
页码:
153-159
栏目:
论著
出版日期:
2013-05-25

文章信息/Info

Title:
CD133 positive U87 glioma stem cell radiosensitivity and DNA double-strand break repair
作者:
李平;  宗天舟;  季晓芹;  陆雪官
215004 苏州, 苏州大学附属第二医院放疗科
Author(s):
LI Ping;  ZONG Tian-zhou;  JI Xiao-qin;  LU Xue-guan
Department of Radiation Oncology, the Second Affiliated Hospital of Soochow University, Suzhou 215004, China
关键词:
神经胶质瘤; 干细胞; 辐射耐受性; DNA修复; 组蛋白类; Rad51重组酶
Keywords:
Glioma; Stem cells; Radiation tolerance; DNA repair; Histones; Rad51 recombinase
DOI:
10.3760/cma.j.issn.1673-4114.2013.03.007
摘要:
目的 探讨CD133+U87人脑胶质瘤干细胞放射敏感性及DNA双链断裂损伤修复的情况。方法 选择人脑胶质瘤U87细胞系,采用免疫流式分选技术分选出CD133+、CD133-细胞;采用克隆形成实验研究细胞的放射敏感性;采用中性单细胞凝胶电泳实验检测4GyX射线垂直照射后不同时间点的DNA双链断裂;采用间接免疫荧光技术检测不同时间点磷酸化组蛋白H2AX(γ-H2AX)荧光灶、Rad51(一种同源重组修复蛋白)荧光灶的表达。结果 假照射条件下,CD133+细胞克隆的形成率明显高于CD133-细胞(t=3.66,P<0.01);CD133+细胞经4 Gy照射后的克隆形成率无明显变化(t=0.71,P>0.05),而CD133-细胞经4 Gy照射后的克隆形成率下降(t=2.91,P<0.05)。4 Gy照射后0.5 h,CD133+、CD133-细胞间尾力矩差异无统计学意义(t=1.44,P>0.05),照射后6、24 h,CD133+细胞尾力距下降程度大于CD133-细胞(t=5.31和8.09,P<0.01);照射后0.5、6h,CD133+、CD133-细胞间γ-H2AX灶的表达率差异均无统计学意义(t=0.12和0.99,P>0.05),照射后24 h,CD133+细胞的γ-H2AX灶的表达率下降程度大于CD133-细胞(t=4.99,P<0.01);照射后0.5 h,CD133+、CD133-细胞间Rad51灶的表达率差异无统计学意义(t=1.12,P>0.05),照射后6、24 h,CD133-细胞的Rad51灶的表达率与CD133+细胞相比明显下降(t=22.88和12.43,P<0.01),而CD133+细胞无明显变化。结论 CD133+U87人脑胶质瘤干细胞具有放射抵抗性,可能与其照射后DNA双链的断裂修复能力较高有关。
Abstract:
Objective To explore the radiosensitivity and DNA double-strand break repair of CD133+U87 glioma stem cell.Methods CD133+ and CD133- cells were isolated from glioma U87 cell lines by flow cytometry sorter system.After irradiated vertically by 4 Gy X-rays,the radiosensitivity of cells was determined by clonogenic assay.The radiation-induced DNA double-strand break repair of CD133+ and CD133- cells was determined by the neutral comet assay,and the expression of phosphorylated histone H2AX (γ-H2AX) and Rad51 foci were measured by immunofluorescence.Results The clone forming rate of CD133+ cells was higher than CD133- cells (t=3.66,P<0.01) with no radiation.The clone forming rate of CD133+ cells irradiated by 4 Gy X-rays has no significant changes compared to that of the non-irradiation cells (t=0.71,P>0.05),but for CD 133 cells,it decreased compared to non-irradiation cells (t=2.91,P<0.05).The tailmoment between CD133+ cells and CD133- cells had no difference at 0.5 h after irradiation (t=1.44,P>0.05); the tailmoment of CD133+ cells was lower than CD133 cells at 6 and 24 h after irradiation,respectively (t=5.31 and 8.09,P<0.01).There was no significant difference in the expression of γ-H2AX foci between CD133+ and CD133- cells at 0.5 and 6 h after irradiation(t=0.12 and 0.99,P>0.05),γ-H2AX foci of CD133+ cells was significantly decreased compared to CD133- cells at 24 h after irradiation (t=4.99,P<0.01).For Rad 51 foci,there was no difference between CD133+ and CD133- cells at 0.5 h after irradiation(t=1.12,P>0.05).The expression of Rad 51 foci of CD133- cells was decreased compared to that of CD133+ cells at 6 and 24 h after irradiation,respectively (t=22.88 and 12.43,P<0.01).And the expression of Rad51 foci of CD133+ cells had no significant changes at 6-24 h after irradiation.Conclusions Glioma stem cells is more radioresistive than glioma non-stem cells.The probable mechanism is that the DNA double-strand break repair capacity of glioma stem cells is more powerful than non-stem cells.

参考文献/References:

[1] Singh SK,Clarke ID,Terasaki M,et al. Identification of a cancer stem cell in human brain tumors. Cancer Res,2003,63(18):5821-5828.
[2] Bao S,Wu Q,Mtrlendon RE,et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature,2006,444(7120):756-760.
[3] 张军宁,朱寿彭,洪承皎. 中性单细胞电泳检测肿瘤细胞放射敏感性. 中华放射医学与防护杂志,2003,23(3):166-168.
[4] Tashiro S,Walter J,Shinohara A,et al. Rad51 accumulation at sites of DNA damage and in postreplicative chromatin. J Cell Biol,2000,150(2):283-291.
[5] Singh SK,Hawkins C,Clarke ID,et al. Identification of human brain tumour initiating cells. Nature,2004,432(7015):396-401.
[6] McCord AM,Jamal M,Williams ES,et al. CD133+ glioblastoma stem-like cells are radiosensitive with a defective DNA damage response compared with established cell iines. Clin Cancer Res,2009,15(16):5145-5153.
[7] Olive PL,Frazer G,Bánath JP. Radiation-induced apoptosis measured in TK6 human B lymphoblast cells using the comet assay. Radiat Res,1993,136(1):130-136.
[8] Pauli TT,Rogakou EP,Yamazaki V,et al,A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol,2000,10(15):886-895.
[9] Celeste A,Petersen S,Romanienko PJ,et al. Genomic instability in mice lacking histone H2AX. Science,2002,296(5569):922-927.
[10] Rogakou EP,Pilch DR,Orr AH,et al. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem,1998,273(10):5858-5868.
[11] Golding SE,Rosenberg E,Khalil A,et al. Double strand break repair by homologous recombination is regulated by cell cycle-independent signaling via ATM in human glioma cells. J Biol Chem,2004,279(15):15402-15410.
[12] Tarsounas M,Davies D,West SC. BRCA2-dependent and independent fonnation of RAD51 nuclear foci. Oncogene,2003,22(8):1115-1123.

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备注/Memo

备注/Memo:
收稿日期:2012-11-26。
基金项目:2011年江苏省"科教兴卫"医学重点人才项目(NO.144);江苏高校优势学科建设工程项目(PAPD)
通讯作者:陆雪官,Email:luxueguanok@yahoo.com.cn
更新日期/Last Update: 1900-01-01