A Novel mRNA-cDNA interference phenomenon for silencing bcl-2 expression in human LNCaP cells.
The templates required for inducing posttranscriptional gene silencing (PTGS) effects have been investigated in human prostate cancer LNCaP cells. Transfection of a mRNA-cDNA hybrid construct was found to result in a relatively long-term interference of specific gene expression. Androgen-stimulated expression of bcl-2 has been reported to increase the tumorigenic and metastatic potentials of human prostate cancer LNCaP cells, as well as their resistance to many apoptotic stimuli. The addition of bcl-2 antisense oligonucleotides, however, restored apoptosis. Our studies demonstrate gene silencing effects of the mRNA-cDNA transfection that is similar to those of PTGS/RNAi in this in vitro prostate cancer cell model. A potential RNA-directed RNA polymerase activity was also detected which is alpha-amanitin-sensitive. These findings indicate that a novel gene silencing system may exist in mammalian cells.
Lin
S L
SL
Department of Pathology, Keck School of Medicine, University of Southern California, HMR-209, 2011 Zonal Avenue, Los Angeles, California, 90033, USA.
Chuong
C M
CM
Ying
S Y
SY
D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D020868:Gene Silencing; D019254:Genes, bcl-2; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger