Details

Autor(en) / Beteiligte

• Yuan, Ya-Wei
• Sun, Ai-Min
• Lui, Ying
• Chen, Long-Hua
• Banerjee, A G

Titel

RNA INTERFERENCE OF ANNEXIN Ⅱ GENE IN PC3 CELLS BY USING SMALL INTERFERENCE RNA SYNTHESIZED WITH IN VITRO TRANSCRIPTION

Ist Teil von

• Chinese medical sciences journal, 2007-03, Vol.22 (1), p.33-37

Ort / Verlag

China: Department of Radiotherapy, Nanfang Hospital of Nanfang Medical University, Guangzhou 510515%Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE 68198-7835 , USA

Erscheinungsjahr

2007

Quelle

MEDLINE

Beschreibungen/Notizen

• Objective To silence annexin Ⅱ gene expression by using small interference RNA （siRNA） in prostate cancer cell line PC3. Methods For in vitro transcription, four sequences of 29-nucleotide DNA template oligonucleotides were designed, and one pair of the sequences were complementary to annexin Ⅱ gene. The other pair was negative control. The 8 nucleotides at the 3＇ end of each oligonucleotide were complementary to the T7 Promoter Primer. The sense and anti-sense siRNA templates were transcribed by T7 RNA polymerase and the resulting RNA transcripts were hybridized to create dsRNA. The siRNA was transfected into prostate cancer cell PC3. For assaying the efficiency of siRNA, confocal microscopy, Northern blotting, and Western blotting were employed to examine the expression of annexin Ⅱ protein and its mRNA. ^3H thymidine was used to measure DNA synthesis. Results The siRNA sequence specific to annexin Ⅱ gene was capable of inhibiting the expression of annexin Ⅱ protein and its mRNA. And cellular DNA synthesis was significantly reduced in siRNA transfected cells. Conclusions The protocol for the synthesis of siRNA by T7 RNA polymerase is feasible. Annexin Ⅱ might be involved in DNA synthesis.