中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2014年
11期
823-826
,共4页
袁晓丽%李涛%黄建鸣%查晓%邓碧芳%郎锦义
袁曉麗%李濤%黃建鳴%查曉%鄧碧芳%郎錦義
원효려%리도%황건명%사효%산벽방%랑금의
食管肿瘤%自噬%氯喹%放射敏感性
食管腫瘤%自噬%氯喹%放射敏感性
식관종류%자서%록규%방사민감성
Esophageal neoplasms%Autophagy%Chloroquine%Radiosensitivity
目的 探讨氯喹对食管癌TE-1细胞系的放射增敏作用及其主要机制.方法 采用MTT法检测不同浓度氯喹对TE-1细胞的生长抑制作用.分别用单纯照射或照射前联合氯喹、照射后联合氯喹作用于TE-1细胞,作用6h后用Western blot测定自噬相关蛋白LC3和Beclin1的表达,用Lyso-Tracker Red DND-99/Hochest 33258进行荧光染色,并用荧光显微镜观察细胞内酸性囊泡(AVOs)的变化.克隆形成实验分析细胞增殖的改变,拟合剂量-生存曲线并计算放射增敏参数.结果 氯喹对TE-1细胞的生长抑制作用呈浓度依赖性.辐射显著诱导TE-1细胞LC3和Beclin1的表达,并促进LC3-Ⅰ转换为LC3-Ⅱ.与单纯照射及照射前加药相比,照射后加药显著降低TE-1细胞内AVOs的荧光强度(F=16.44,P＜0.05)和克隆形成率,照射前加药和照射后加药的SERD0分别为1.037和1.439(t =8.30,P＜0.05).结论 氯喹能增加食管鳞癌TE-1细胞的放射敏感性,其机制可能与抑制自噬作用相关.
目的 探討氯喹對食管癌TE-1細胞繫的放射增敏作用及其主要機製.方法 採用MTT法檢測不同濃度氯喹對TE-1細胞的生長抑製作用.分彆用單純照射或照射前聯閤氯喹、照射後聯閤氯喹作用于TE-1細胞,作用6h後用Western blot測定自噬相關蛋白LC3和Beclin1的錶達,用Lyso-Tracker Red DND-99/Hochest 33258進行熒光染色,併用熒光顯微鏡觀察細胞內痠性囊泡(AVOs)的變化.剋隆形成實驗分析細胞增殖的改變,擬閤劑量-生存麯線併計算放射增敏參數.結果 氯喹對TE-1細胞的生長抑製作用呈濃度依賴性.輻射顯著誘導TE-1細胞LC3和Beclin1的錶達,併促進LC3-Ⅰ轉換為LC3-Ⅱ.與單純照射及照射前加藥相比,照射後加藥顯著降低TE-1細胞內AVOs的熒光彊度(F=16.44,P＜0.05)和剋隆形成率,照射前加藥和照射後加藥的SERD0分彆為1.037和1.439(t =8.30,P＜0.05).結論 氯喹能增加食管鱗癌TE-1細胞的放射敏感性,其機製可能與抑製自噬作用相關.
목적 탐토록규대식관암TE-1세포계적방사증민작용급기주요궤제.방법 채용MTT법검측불동농도록규대TE-1세포적생장억제작용.분별용단순조사혹조사전연합록규、조사후연합록규작용우TE-1세포,작용6h후용Western blot측정자서상관단백LC3화Beclin1적표체,용Lyso-Tracker Red DND-99/Hochest 33258진행형광염색,병용형광현미경관찰세포내산성낭포(AVOs)적변화.극륭형성실험분석세포증식적개변,의합제량-생존곡선병계산방사증민삼수.결과 록규대TE-1세포적생장억제작용정농도의뢰성.복사현저유도TE-1세포LC3화Beclin1적표체,병촉진LC3-Ⅰ전환위LC3-Ⅱ.여단순조사급조사전가약상비,조사후가약현저강저TE-1세포내AVOs적형광강도(F=16.44,P＜0.05)화극륭형성솔,조사전가약화조사후가약적SERD0분별위1.037화1.439(t =8.30,P＜0.05).결론 록규능증가식관린암TE-1세포적방사민감성,기궤제가능여억제자서작용상관.
Objective To investigate the possibility of chloroquine radiosensitization of esophageal cancer cell line TE-1 and its further mechanism.Methods Effect of chloroquine on cell viability of TE-1 cells was determined by MTT method.Expression of LC3,Beclin-1 and formation of acidic vesicular organelles (AVOs) were determined by Western blot,and fluorescence staining with Lyso-Tracker Red DND-99,respectively.Clonogenic survival of TE-1 cells was examined by clonogenic forming assay.Results Chloroquine showed dose-dependent inhibition of TE-1 cell growth,and its values of IC50 and IC10 were (72.33 ± 5.28) and (15.42 ± 3.33) μmol/L,respectively.The expression of Beclin-1 and LC3-Ⅱ/Ⅰ markedly increased in irradiated TE-1 cells.The addition of chloroquine with IC10concentration significantly reduced the fluorescence and intensity of AVOs accumulation in the cytoplasm of TE-1 cells.Clonogenic survival fraction decreased obviously,in TE-1 cells with addition of chloroquine after radiation and the value of SERD0 was 1.439.Conclusions Chloroquine could radiosensitize esophageal cancer cells by blocking autophagy-lysosomal pathway and be used as a potential radiosensitizing strategy.
