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| Survivin基因RNAi真核表达载体对HL-60细胞增殖及凋亡的影响 | |
| 引用本文： | 王琰,孙玲,赵国强,王业生. Survivin基因RNAi真核表达载体对HL-60细胞增殖及凋亡的影响[J]. 中国现代医学杂志, 2008, 18(4): 454-457 |
| 作者姓名： | 王琰 孙玲 赵国强 王业生 |
| 作者单位： | 1. 郑州大学第五附属医院,血液科,河南,郑州,450052 2. 郑州大学第一附属医院,血液科,河南,郑州,450052 3. 郑州大学基础医学院,免疫教研室,河南,郑州,450052 4. 郑州大学第一附属医院,血液科,河南,郑州450052 |
| 摘 要： | 目的 构建Survivin的shRNA表达质粒并将其导入白血病细胞株HL-60细胞以探讨PNA干扰对HL-60细胞增殖及凋亡的影响.方法 设计、合成两对针对survivin的shRNA序列,连接到带有人U6启动子的裁体质粒pSINsi-Hu6中,将构建重组质粒命名为pSIN/shRNA1和pSIN/shRNA2.pSIN/shRNA1转染HL60细胞,应用四唑盐(MTT)比色法观察细胞增殖情况;流式细胞仪检测细胞凋亡.结果 酶切分析和测序证实pSIN/shRNA1和pSIN/shRNA2构建成功.MTT测定显示转染重组质粒pSIN/shRNA1入HL-60细胞48、72和96 h后细胞增殖明显受到抑制,分别与阴性对照组、空白对照组组比较,差异有显著性(P＜0.05).流式细胞仪分析pSIN/shRNA1组胞凋亡率达(20.21±0.75)%,明显高于阴性对照组的(2.58±0.48)%和空白对照组的(1.26±0.30)%.结论 成功地构建了survivin基因RNAi真核表达栽体,且pSIN/shRNA1可序列特异性地抑制HL-60细胞的增殖并诱导其凋亡,为白血病的基因治疗奠定基础. |
| 关 键 词： | survivin基因 RNA干扰 短发夹状RNA 白血病 HL60细胞 Survivin 基因治疗 RNAi 真核表达载体 细胞增殖 凋亡率 影响 cells apoptosis proliferation expression vector 细胞的增殖 序列特异性 栽体 流式细胞仪分析 差异 比较 对照组 阴性 显示 |
| 文章编号： | 1005-8982（2008）04-0454-04 |
| 收稿时间： | 2007-08-16 |
| 修稿时间： | 2007-08-16 |
Effect of survivin expression vector on proliferation and apoptosis of HL-60 cells by RNAi |
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| WANG Yan,SUN Ling,ZHAO Guo-qiang,WANG Ye-sheng. Effect of survivin expression vector on proliferation and apoptosis of HL-60 cells by RNAi[J]. China Journal of Modern Medicine, 2008, 18(4): 454-457 | |
| Authors: | WANG Yan SUN Ling ZHAO Guo-qiang WANG Ye-sheng |
| Affiliation: | WANG Yan1,SUN Ling1,ZHAO Guo-qiang2,WANG Ye-sheng1 (1.Department of Hematology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,P.R.China,2.Department of Hematology,the First Affiliated Hospital of Zhengzhou University,3.Department of Microbiology , Immunology,College of Basic Medical Sciences,Zhengzhou University,P.R.China ) |
| Abstract: | Objective To construct the short hairpin RNA (shRNA) expression vector of survivin and investigate its effect on the proliferation and the apoptosis of leukemia cell line HL-60. Methods Two pairs of oligonucleotide sequences targeted at human surviving mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pSINsi-Hu6 plasmid. After being identified by restriction enzyme digestion and sequencing,the recombinant plasmid pSIN/shRNA1 were transfected into HL-60 cells. The... |
| Keywords: | survivin gene RNA interferenee short hairpin RNA leukemia HL-60 eeU |
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