Downregulation of bcl-2 expression in lymphoma cells by bcl-2 ARE-targeted modified, synthetic ribozyme.
Synthetic ribozymes are catalytic RNA molecules designed to inhibit gene expression by cleaving specific mRNA sequences. We investigated the potential of synthetic ribozymes to inhibit bcl-2 expression in apoptosis defective bcl-2 overexpressing tumors. A chemically stabilized hammerhead ribozyme has been targeted to the A+U-rich regulative element of bcl-2 mRNA that is involved in bcl-2 gene switch-off during apoptosis. The design of the ribozyme was based on the results of probing accessibility of the RNA target in cellular extracts with antisense DNA. The ribozyme was lipotransfected to a bcl-2 overexpressing human lymphoma cell line (Raji). The cellular uptake of this ribozyme resulted in a marked reduction of both bcl-2 mRNA and BCL-2 protein levels and dramatically increased cellular death by apoptosis. Our results suggest a potential therapeutic application of such ribozyme for the treatment of bcl-2 overexpressing tumors.
Luzi
Ettore
E
Department of Experimental Pathology and Oncology, University of Florence, Italy. et.car@dada.it
Papucci
Laura
L
Schiavone
Nicola
N
Donnini
Martino
M
Lapucci
Andrea
A
Tempestini
Alessio
A
Witort
Ewa
E
Nicolin
Angelo
A
Capaccioli
Sergio
S
D017209:Apoptosis; D001483:Base Sequence; D002051:Burkitt Lymphoma / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005813:Genes, Synthetic; D006801:Humans; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid
D020319:Oligodeoxyribonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D016337:RNA, Catalytic; D012333:RNA, Messenger; D038621:Regulatory Sequences, Ribonucleic Acid; C427204:hammerhead ribozyme