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Inhibition of Liposome Transfection siRNA on HDAC1 Expression in Mouse Renal Tubular Epithelial Cells

YANG Yang1,2, WU Guo-juan2, LI Pei-feng1   

  1. 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
  • Received:2012-02-07 Online:2012-08-20 Published:2012-08-24

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 LipofectamineTM 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.

Key words: HDAC1-siRNA; RTECs; FAM-siRNA; MTT

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