Induction of apoptosis in HeLa cells with siRNA expression vector targeted against bcl-2.
RNA interference (RNAi) is the process by which double-stranded RNA (dsRNA) directs sequence-specific gene silencing in animal and plant cells. In mammalian cells, 21- or 22-nucleotide (nt) RNAs with 2-nt 3' overhangs small inhibitory RNAs (siRNAs) exhibit an RNAi effect. Very recently, we and others have developed a vector-based siRNA expression system that can induce RNAi in mammalian cells. In this report, to apply this system to oncogene therapy, we tried to suppress the expression of the bcl-2 gene, which is known as a key molecule in the regulation of apoptosis or programmed cell death, by using the siRNA expression system. Western blotting analysis revealed that this siRNA expression vector against bcl-2 suppressed the expression of the bcl-2 protein. Furthermore, HeLa cells which were transiently transfected with the siRNA expression vector against bcl-2 and were subsequently treated with doxorubicin efficiently underwent apoptosis, concomitant with the repression of the bcl-2 gene. These results demonstrate that the siRNA expression vector against bcl-2 has a potential as therapeutic agent for a variety of cancers by down-regulating bcl-2 gene expression.
Futami
Takashi
T
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, Hongo, Tokyo 113-8656, Japan.
Miyagishi
Makoto
M
Seki
Minoru
M
Taira
Kazunari
K
D017209:Apoptosis / Q000235:genetics*; D019254:Genes, bcl-2; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
D034741:RNA, Small Interfering