白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2011年
6期
357-361
,共5页
张旭艳%卓家才%杜新%李明%陶小梅%史敦云%楼瑾%张琼丽
張旭豔%卓傢纔%杜新%李明%陶小梅%史敦雲%樓瑾%張瓊麗
장욱염%탁가재%두신%리명%도소매%사돈운%루근%장경려
RNA,小分子干扰%DNA拓扑异构酶类,Ⅱ型%砷酸盐类%K562/AS2细胞
RNA,小分子榦擾%DNA拓撲異構酶類,Ⅱ型%砷痠鹽類%K562/AS2細胞
RNA,소분자간우%DNA탁복이구매류,Ⅱ형%신산염류%K562/AS2세포
RNA,small hairpin%DNA topoisomerases,tope Ⅱ%Arsenates%K562/AS2
目的 研究小干扰RNA片段(shRNA)对三氧化二砷(ATO)耐药的白血病细胞株K562/AS2细胞的Topo Ⅱα、TopoⅡβ基因表达及其功能的影响.方法 设计并合成针对Topo Ⅱα和TopoⅡβ基因序列的shRNA各3对,在脂质体的介导下转染K562/AS2细胞;用荧光实时定量聚合酶链反应(PCR)分析Topo Ⅱα、TopoⅡβmRNA的表达水平;流式细胞术检测Topo Ⅱα、TopoⅡβ蛋白表达.结果 针对Topo Ⅱα-shRNA、TopoⅡβ的shRNA作用于K562/AS2细胞24 h后,Topo ⅡαmRNA水平和蛋白水平最大下调为(78.22±0.01)%、(31.17±1.27)%(P<0.05),TopoⅡβmRNA水平和蛋白水平最大下调为(57.36 ±0.01)%、(23.98 ± 1.22)%(P<0.05).结论 转染24 h后针对TopoⅡ的shRNA可抑制对ATO耐药的白血病细胞株K562/AS2细胞TopoⅡ基因的表达.
目的 研究小榦擾RNA片段(shRNA)對三氧化二砷(ATO)耐藥的白血病細胞株K562/AS2細胞的Topo Ⅱα、TopoⅡβ基因錶達及其功能的影響.方法 設計併閤成針對Topo Ⅱα和TopoⅡβ基因序列的shRNA各3對,在脂質體的介導下轉染K562/AS2細胞;用熒光實時定量聚閤酶鏈反應(PCR)分析Topo Ⅱα、TopoⅡβmRNA的錶達水平;流式細胞術檢測Topo Ⅱα、TopoⅡβ蛋白錶達.結果 針對Topo Ⅱα-shRNA、TopoⅡβ的shRNA作用于K562/AS2細胞24 h後,Topo ⅡαmRNA水平和蛋白水平最大下調為(78.22±0.01)%、(31.17±1.27)%(P<0.05),TopoⅡβmRNA水平和蛋白水平最大下調為(57.36 ±0.01)%、(23.98 ± 1.22)%(P<0.05).結論 轉染24 h後針對TopoⅡ的shRNA可抑製對ATO耐藥的白血病細胞株K562/AS2細胞TopoⅡ基因的錶達.
목적 연구소간우RNA편단(shRNA)대삼양화이신(ATO)내약적백혈병세포주K562/AS2세포적Topo Ⅱα、TopoⅡβ기인표체급기공능적영향.방법 설계병합성침대Topo Ⅱα화TopoⅡβ기인서렬적shRNA각3대,재지질체적개도하전염K562/AS2세포;용형광실시정량취합매련반응(PCR)분석Topo Ⅱα、TopoⅡβmRNA적표체수평;류식세포술검측Topo Ⅱα、TopoⅡβ단백표체.결과 침대Topo Ⅱα-shRNA、TopoⅡβ적shRNA작용우K562/AS2세포24 h후,Topo ⅡαmRNA수평화단백수평최대하조위(78.22±0.01)%、(31.17±1.27)%(P<0.05),TopoⅡβmRNA수평화단백수평최대하조위(57.36 ±0.01)%、(23.98 ± 1.22)%(P<0.05).결론 전염24 h후침대TopoⅡ적shRNA가억제대ATO내약적백혈병세포주K562/AS2세포TopoⅡ기인적표체.
Objective To investigate the reversal effect of Topo Ⅱα-shRNA and Topo Ⅱβ-shRNA on Topo Ⅱ gene in K562/AS2 cells. Methods Three pieces of Topo Ⅱα-shRNA and three pieces of Topo Ⅱβ-shRNA were designed,synthesized and transfected into K562/AS2 cells by liposome. Expression level of Topo Ⅱα and Topo Ⅱβ mRNA were determined by real time fluorescence quantitative PCR. The expressions of Topo Ⅱα and Topo Ⅱβ protein were assayed with flow cytometer. Results After treated with Topo Ⅱα-shRNA or Topo Ⅱβ-shRN A for 24 hours,the expression level of Topo Ⅱα mRNA and Topo Ⅱβ mRNA protein in K562/AS2 cells decreased at most (78.22±0.01) %,(31.17±1.27) % (P <0.05),and (57.36±0.01)%,(23.98±1.22) % (P <0.05) respectively. Conclusion The expression of Topo Ⅱ gene can be down-regulated after infected with Topo Ⅱ-shRNA in K562/ AS2 cell line.
