| 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） |
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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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