Hammerhead ribozyme specifically inhibits mutant K-ras mRNA of human pancreatic cancer cells.
We have evaluated the efficacy of an anti-K-ras hammerhead ribozyme targeted against GUU-mutated codon 12 of the K-ras gene in a cell-free system as well as in cultured pancreatic carcinoma cell lines. In the cell-free system, the anti-K-ras ribozyme specifically cleaved K-ras RNA with GUU-mutation at codon 12, but not other triplet sequences at codon 12 of K-ras RNA. In the cell culture system, the anti-K-ras ribozyme significantly reduced K-ras mRNA level (GUU-mutated codon 12) in Capan-1 pancreatic carcinoma cells, but less significantly suppressed K-ras mRNA in Capan-2 (GUU/GGU heterozygous-mutation at codon 12) or MIA PaCa-2 (UGU-mutated codon 12) pancreatic carcinoma cells. The ribozyme inhibited proliferation of transfected Capan-1 cells. These results suggest that this ribozyme selectively recognizes single-base mutation of K-ras mRNA and is able to reverse the malignant phenotype in human pancreatic carcinoma cells.
Tsuchida
T
T
Department of Pathology, Tokai University School of Medicine, Isehara, Japan.
Kijima
H
H
Oshika
Y
Y
Tokunaga
T
T
Abe
Y
Y
Yamazaki
H
H
Tamaoki
N
N
Ueyama
Y
Y
Scanlon
K J
KJ
Nakamura
M
M
D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002277:Carcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002474:Cell-Free System; D015536:Down-Regulation / Q000235:genetics; D011905:Genes, ras; D006131:Growth Inhibitors / Q000502:physiology; D006801:Humans; D006868:Hydrolysis; D016297:Mutagenesis, Site-Directed; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D016337:RNA, Catalytic / Q000502:physiology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D014407:Tumor Cells, Cultured
D006131:Growth Inhibitors; D016337:RNA, Catalytic; D012333:RNA, Messenger