文章摘要
何敏毅,孙学刚,文 戈,赵 亮.X线照射对宫颈癌HeLa细胞醛缩酶A表达和定位的影响[J].南京医科大学学报,2010,(5):617~620
X线照射对宫颈癌HeLa细胞醛缩酶A表达和定位的影响
Effects of X-ray irradiation on expression and localization of aldolase A in HeLa cells
投稿时间:2009-07-13  
DOI:10.7655
中文关键词: 醛缩酶A  X线  HeLa细胞
英文关键词: aldolase A  X ray  HeLa cells
基金项目:广东省自然科学基金面上项目(06024430);广东省社会发展领域科技计划项目(83007)
作者单位
何敏毅 南方医科大学珠江医院器官移植科,广东 广州 510515 
孙学刚 南方医科大学中医药学院中医症候实验室,广东 广州 510515 
文 戈 南方医科大学南方医院影像中心,广东 广州 510515 
赵 亮 南方医科大学教育部和广东省共建重大疾病转录组学及功能蛋白质组学重点实验室,广东 广州 510515 
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中文摘要:
      目的:观察宫颈癌HeLa细胞转染醛缩酶A(aldolase A,ALDOA) 后,在X线作用下ALDOA在细胞内定位的改变,以及其对细胞存活率的影响?方法:克隆出ALDOA全长编码序列,构建真核表达质粒pECFP-C1-ALDOA,实验组通过脂质体介导将 pECFP-C1-ALDOA转染HeLa 细胞,设载体pECFP-C1作为阴性对照,各组均给予X线照射,荧光显微镜观察照射前后HeLa细胞中pECFP-C1-ALDOA和pECFP-C1表达及定位的变化,用XTT法检测细胞存活率的改变?结果:双酶切和DNA测序证明所构建质粒正确;荧光显微镜下观察HeLa细胞内 pECFP-C1-ALDOA主要分布于胞浆,pECFP-C1为全细胞均匀分布;经X线照射24 h后胞浆中的pECFP-C1-ALDOA逐渐转至胞核,而pECFP-C1在细胞内的定位则不发生明显改变;经XTT法检测,发现X线照射后,转染pECFP-C1-ALDOA质粒的细胞存活率明显高于转染pECFP-C1的对照组细胞?结论:抑制ALDOA的表达,可使X线更易诱导宫颈癌细胞的死亡,因此其可能作为宫颈癌放射治疗的新靶点?
英文摘要:
      Objective:To study the change of the localization of aldolase A (ALDOA) in Hela cells before and after X ray exposure,and impact of the treatment on cell survival rate. Methods:The whole encoding sequence of ALDOA was cloned into the eukaryotic expressing pECFP-C1-ALDOA plasmid,and then transfected into Hela cells in the experimental group. Meanwhile,the pECFP-C1 vector was transfected into Hela cells as a control. The expression and localization of pECFP-C1-ALDOA and pECFP-C1 were investigated by fluorescent microscopy before and after X ray exposure,and the cell survival rate was detected by XTT analysis. Results:The constructed plasmid was verified by double enzyme digestion and DNA sequencing;the fluorescent microscopy revealed that pECFP-C1-ALDOA located predominantly in cell endochylema while pECFP-C1 dispersed equally in the whole cell;pECFP-C1-ALDOA located into cell nucleus 24 hours after X ray exposure,however,the localization of pECFP-C1 did not change significantly;XTT analysis revealed that the survival rate of the pECFP-C1-ALDOA-transfected Hela cells was much higher than untransfected cells after X ray exposure. Conclusion:Inhabiting the expression of ALDOA may increase Hela cell death induced by X-ray,so ALDOA could be used as a new target for cancer radiation therapy.
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