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| 脂质体瞬时转染siRNA抑制肾小管上皮细胞去乙酰化蛋白酶1的表达 | |
| 引用本文： | 杨洋,吴国娟,李培锋.脂质体瞬时转染siRNA抑制肾小管上皮细胞去乙酰化蛋白酶1的表达[J].中国畜牧兽医,2012,39(8):4-10. |
| 作者姓名： | 杨洋 吴国娟 李培锋 |
| 作者单位： | 1. 内蒙古农业大学兽医学院,内蒙古呼和浩特 010018; 2. 北京农学院动物科学技术学院,北京 102206 |
| 基金项目： | 国家自然科学基金资助项目，北京市自然科学基金资助 |
| 摘 要： | 建立有效的脂质体瞬时转染法,将小干扰RNA(siRNA)转染至小鼠肾小管上皮细胞(renal tubular epithelial cells,RTECs),从而抑制其乙酰化组蛋白酶1(histone deacetylases 1,HDAC1)的表达。使用阳离子脂质体法瞬时转染小鼠肾小管上皮细胞,用荧光标记siRNA(FAM-siRNA)筛选转染比例,实时荧光定量PCR(Real-time PCR)确定最优条件并检测不同位点(374,525和822)HDAC1-siRNA的抑制效果,噻唑蓝(MTT)法检测不同时间点HDAC1-siRNA对RTECs增殖的影响,并设立曲古霉素处置组作为试验对照。荧光表达和Real-time PCR结果显示30 pmol siRNA∶1.5 μL LipofectamineTM 2000为最佳转染条件,HDAC1-374的干扰效果最明显。MTT检测结果显示,HDAC1-374与TSA一致,证实HDAC1-siRNA能有效抑制RTECs的增殖。本试验利用脂质体瞬时转染siRNA法成功抑制了RTECs中HDAC1的表达。 |
| 关 键 词： | HDAC1-siRNA RTECs FAM-siRNA MTT |
| 收稿时间： | 2012-02-07 |
Inhibition of Liposome Transfection siRNA on HDAC1 Expression in Mouse Renal Tubular Epithelial Cells |
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| YANG Yang , WU Guo-juan , LI Pei-feng.Inhibition of Liposome Transfection siRNA on HDAC1 Expression in Mouse Renal Tubular Epithelial Cells[J].China Animal Husbandry & Veterinary Medicine,2012,39(8):4-10. | |
| Authors: | YANG Yang WU Guo-juan LI Pei-feng |
| Institution: | 1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China |
| Abstract: | To transfect histone deacetylases 1(HDAC1)-small interferencing RNA(siRNA) into mouse renal tubular epithelial cells(RTECs) by liposome transfection and to investigate its inhibitory effect on HDAC1.It was transient into RTECs by liposome transfection.Fluorescently labeled siRNA(the FAM-siRNA) to filter the proportion of transfected.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) to determine the optimal conditions and testing the various sites(374,525 and 822) HDAC1-siRNA inhibition effect,MTT method to detect the influence of the HDAC1-siRNA on suppression RTECs proliferation in different time points,and trichostatin A(TSA) disposal group as a test control.Expression of fluorescence and Real-time PCR results showed that 30 pmol siRNA∶ 1.5 mL Lipofectamine TM 2000 were optimal transfection conditions and HDAC1-374 had the most obvious interference effect.The MTT testing results of HDAC1-374 and TSA were consistent: HDAC1-siRNA could effectively restrain RTECs proliferation.The test using liposomes transiently transfected siRNA method,successfully inhibition the expression of HDAC1 in RTECs. |
| Keywords: | HDAC1-siRNA RTECs FAM-siRNA MTT |
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