[1]李庆霞,王娟,赵文清,等.RNA干扰survivin基因增强SKBr-3细胞对多柔吡星敏感性的体外研究[J].第三军医大学学报,2010,32(11):1158-1161.
 Li Qingxia,Wang Juan,Zhao Wenqing,et al.RNAinterfered survivin gene increases sensitivity of SKBr-3 cells to doxorubicin: an in vitro study[J].J Third Mil Med Univ,2010,32(11):1158-1161.
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RNA干扰survivin基因增强SKBr-3细胞对多柔吡星敏感性的体外研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第11期
页码:
1158-1161
栏目:
论著
出版日期:
2010-06-15

文章信息/Info

Title:
RNAinterfered survivin gene increases sensitivity of SKBr-3 cells to doxorubicin: an in vitro study
作者:
李庆霞王娟赵文清刘家云黄红艳张明杨安钢
河北省人民医院肿瘤一科;第四军医大学:附属西京医院检验科,免疫学教研室;军事医学科学院附属307医院肿瘤三科
Author(s):
Li Qingxia Wang Juan Zhao Wenqing Liu Jiayun Huang Hongyan Zhang Ming Yang Angang
First Department of Oncology, People’s  Hospital of Hebei Province, Shijiazhuang, Hebei Province, 050051;Department of Clinical Laboratory, Xijing Hospital, Department of Immunology, College of Basic Medical Sciences, Fourth Military Medical University, Xi′an, Shaanxi Province,710032;Third Department of Oncology, No. 307 Hospital of PLA, Beijing, 100071, China
关键词:
RNA干扰survivinSKBr-3细胞药物敏感性
Keywords:
RNA interference survivin SKBr-3 cells chemosensitivity
分类号:
R73-362;R730.53;R737.9
文献标志码:
A
摘要:
目的    探讨抑制survivin基因的表达对人乳腺癌SKBr-3细胞化疗敏感性的影响。    方法    构建针对survivin基因序列特异性shRNA的表达载体,脂质体转染SKBr-3,G418筛选阳性克隆。实验分为RNA干扰组(S1&P),脂质体对照组(H1&P)和空白对照组。光镜观察转染后细胞的形态变化;0.5 μg/ml 多柔吡星作用于转染后的细胞,电镜观察细胞形态学变化;TUNEL染色、Annexin V荧光标记检测细胞凋亡,MTT法检测细胞增殖的抑制率。    结果    成功构建具有新霉素抗性的靶向survivin的序列特异性shRNA表达载体,基因测序完全正确;转染后干扰组细胞均发生细胞凋亡的形态学改变;低剂量多柔吡星诱导后,TUNEL染色结果,S1&P组凋亡细胞比例(23.6±4.8)%明显高于H1&P组(4.1±1.1) %和空白对照组(3.2±1.4) %(P<0.05),表明转染了RNA干涉载体的SKBr-3细胞对低剂量多柔吡星的敏感性显著增加;Annexin-V染色结果:空白对照组凋亡比例为(17.43±3.12)%,H1&P组为(20.51±4.67)%,S1&P组为(68.76±8.49)%,后者与前两者比较均有统计学差异(P<0.05)。干扰组细胞凋亡率、细胞增殖的抑制率均明显增加,差异有统计学意义(P<0.05)。    结论    体外实验证实抑制survivin基因的表达可提高乳腺癌SKBr-3细胞对多柔吡星的敏感性。
Abstract:
Objective    To study the effect of RNA-interfered survivin gene on chemosensitivity of breast cancer cell line SKBr-3 to doxorubicin.    Methods    A shRNA expression vector,ΔpcDNA3-H1-S1, was constructed according to the specificity of survivin gene. SKBr-3 cells were transfected with liposomeencapsulated plasmids and positive clones were screened with G418. SKBr-3 cells were divided into RNA interference group, liposome control group and blank control group. Change of morphology in cells transfected with 0.5 μg/ml doxorubicin was observed under an optical or electron microscope. Cell apoptosis was detected by TUNEL assay with Annexin-Ⅴ  fluorescence staining, and inhibiting rate of cell proliferation was detected by MTT assay.     Results    The survivin gene sequence specific shRNA expression vector resistant to neomycin was successfully constructed, which was confirmed by DNA sequencing. The morphologic change of apoptotic cells was observed in RNA interference group. The rate of apoptotic cells was significantly higher in RNA interference group than in liposome control group and blank control group [(23.6±4.8)% vs (4.1±1.1)% and (3.2±1.4)%] after TUNEL staining (P<0.05), indicating that the sensitivity of SKBr-3 cells to doxorubicin was increased in RNA-interfered vector. The rate of apoptotic cells was significantly lower in liposome control group and blank control group than in RNA interference group [(17.43±3.12)% and (20.51±4.67)% vs (68.76±8.49)%] after Annexin-V staining (P<0.05).     Conclusion    The chemosensitivity of  SKBr-3 cells to doxorubicin can be significantly increased by inhibiting the expression of survivin gene with RNA interference technology.

参考文献/References:

李庆霞,王娟,赵文清,等.RNA干扰survivin基因增强SKBr-3细胞对多柔吡星敏感性的体外研究[J].第三军医大学学报,2010,32(11):1158-1161.

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更新日期/Last Update: 2010-06-04