国际放射医学核医学杂志
國際放射醫學覈醫學雜誌
국제방사의학핵의학잡지
INTERNATIONAL JOURNAL OF RADIATION MEDICINE AND NUCLEAR MEDICINE
2013年
3期
153-159
,共7页
李平%宗天舟%季晓芹%陆雪官
李平%宗天舟%季曉芹%陸雪官
리평%종천주%계효근%륙설관
神经胶质瘤%干细胞%辐射耐受性%DNA修复%组蛋白类%Rad51重组酶
神經膠質瘤%榦細胞%輻射耐受性%DNA脩複%組蛋白類%Rad51重組酶
신경효질류%간세포%복사내수성%DNA수복%조단백류%Rad51중조매
Glioma%Stem cells%Radiation tolerance%DNA repair%Histones%Rad51 recombinase
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.
目的 探討CD133+ U87人腦膠質瘤榦細胞放射敏感性及DNA雙鏈斷裂損傷脩複的情況.方法 選擇人腦膠質瘤U87細胞繫,採用免疫流式分選技術分選齣CD133+、CD133-細胞；採用剋隆形成實驗研究細胞的放射敏感性；採用中性單細胞凝膠電泳實驗檢測4GyX射線垂直照射後不同時間點的DNA雙鏈斷裂；採用間接免疫熒光技術檢測不同時間點燐痠化組蛋白H2AX(γ-H2AX)熒光竈、Rad51(一種同源重組脩複蛋白)熒光竈的錶達.結果 假照射條件下,CD133+細胞剋隆的形成率明顯高于CD133-細胞(t=3.66,P＜0.01)；CD133+細胞經4 Gy照射後的剋隆形成率無明顯變化(t=0.71,P＞0.05),而CD133-細胞經4 Gy照射後的剋隆形成率下降(t=2.91,P＜0.05).4 Gy照射後0.5 h,CD133+、CD133-細胞間尾力矩差異無統計學意義(t=1.44,P＞0.05),照射後6、24 h,CD133+細胞尾力距下降程度大于CD133-細胞(t=5.31和8.09,P＜0.01)；照射後0.5、6h,CD133+、CD133-細胞間γ-H2AX竈的錶達率差異均無統計學意義(t=0.12和0.99,P＞0.05),照射後24 h,CD133+細胞的γ-H2AX竈的錶達率下降程度大于CD133-細胞(t=4.99,P＜0.01)；照射後0.5 h,CD133+、CD133-細胞間Rad51竈的錶達率差異無統計學意義(t=1.12,P＞0.05),照射後6、24 h,CD133-細胞的Rad51竈的錶達率與CD133+細胞相比明顯下降(t=22.88和12.43,P＜0.01),而CD133+細胞無明顯變化.結論 CD133+U87人腦膠質瘤榦細胞具有放射牴抗性,可能與其照射後DNA雙鏈的斷裂脩複能力較高有關.
목적 탐토CD133+ U87인뇌효질류간세포방사민감성급DNA쌍련단렬손상수복적정황.방법 선택인뇌효질류U87세포계,채용면역류식분선기술분선출CD133+、CD133-세포；채용극륭형성실험연구세포적방사민감성；채용중성단세포응효전영실험검측4GyX사선수직조사후불동시간점적DNA쌍련단렬；채용간접면역형광기술검측불동시간점린산화조단백H2AX(γ-H2AX)형광조、Rad51(일충동원중조수복단백)형광조적표체.결과 가조사조건하,CD133+세포극륭적형성솔명현고우CD133-세포(t=3.66,P＜0.01)；CD133+세포경4 Gy조사후적극륭형성솔무명현변화(t=0.71,P＞0.05),이CD133-세포경4 Gy조사후적극륭형성솔하강(t=2.91,P＜0.05).4 Gy조사후0.5 h,CD133+、CD133-세포간미력구차이무통계학의의(t=1.44,P＞0.05),조사후6、24 h,CD133+세포미력거하강정도대우CD133-세포(t=5.31화8.09,P＜0.01)；조사후0.5、6h,CD133+、CD133-세포간γ-H2AX조적표체솔차이균무통계학의의(t=0.12화0.99,P＞0.05),조사후24 h,CD133+세포적γ-H2AX조적표체솔하강정도대우CD133-세포(t=4.99,P＜0.01)；조사후0.5 h,CD133+、CD133-세포간Rad51조적표체솔차이무통계학의의(t=1.12,P＞0.05),조사후6、24 h,CD133-세포적Rad51조적표체솔여CD133+세포상비명현하강(t=22.88화12.43,P＜0.01),이CD133+세포무명현변화.결론 CD133+U87인뇌효질류간세포구유방사저항성,가능여기조사후DNA쌍련적단렬수복능력교고유관.
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-ceils 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 CD 133+ cells and CD 133-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.
