Abstract
To construct the retroviral vector of p(125FAK) specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapy in vitro. A hammerhead ribozyme DNA targeting p(125FAK) mRNA from nt 1010 to nt 1032 was synthesized and recombined into the retroviral vector pLXSN forming pLRZXSN recon. Using the lipofectin-mediated DNA transfection technique, pLRZXSN was introduced into BGC-823 cells. The effects of ribozyme on the growth of BGC-823 cells and apoptosis were studied by cell colony assay, flow cytometry (FCM), reverse transcriptase-polymerase chain reaction (RT-PCR), detection of DNA fragmentation and electron microscopy. The number of BGC-823 cell colonies was inhibited by 56% after the cells were treated for 48 h. The cell proliferation was inhibited effectively by p(125FAK) ribozyme and the inhibitory effect depended on the concentration and the time of incubation. The expression of p(125FAK) mRNA and protein P(125FAK) decreased sharply in BGC-823 cells treated with p(125FAK) ribozyme. The characteristics of apoptosis, namely sub-G1 peak, DNA fragmentation and morphological changes, were revealed in BGC-823 cells treated with p(125FAK) ribozyme. p(125FAK) ribozyme decreases p(125FAK) gene expression and...Continue Reading
References
Jan 1, 1992·Annual Review of Biochemistry·R H Symons
Aug 18, 1988·Nature·J Haseloff, W L Gerlach
Nov 1, 1994·British Journal of Haematology·Y Z ChenZ C Han
Feb 1, 1993·The Journal of Cell Biology·R L Juliano, S Haskill
May 31, 2002·Oncogene·Orlie KahanaIlana Yron
Sep 7, 2002·Antioxidants & Redox Signaling·Tadashi KasaharaYoshiko Sonoda
Mar 28, 2003·The Biochemical Journal·Lucia BevigliaWilliam G Cance
Aug 6, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kyung-Sook ShinSeong-Wook Lee
Sep 11, 2004·Biochemical and Biophysical Research Communications·Shaker AbuharbeidAchim Aigner
Nov 17, 2004·Current Pharmaceutical Biotechnology·Kevin J Scanlon
Nov 24, 2004·Chemistry & Biology·Dong-Eun Kim, Gerald F Joyce
Feb 3, 2005·Nature Reviews. Molecular Cell Biology·Satyajit K MitraDavid D Schlaepfer
Mar 18, 2005·The Journal of Gene Medicine·Yu-Hu SongYou-Xin Jin
Mar 25, 2005·Oligonucleotides·Shenxian QianYun Yen
Apr 27, 2005·Biochemical and Biophysical Research Communications·Jeeva VaradarajuluChristof R Hauck
May 5, 2005·Nucleic Acids Research·Sylvia TobéMarie-Christine Maurel
Jun 3, 2005·Cancer Research·Maroesja J van NimwegenBob van de Water