Ribozyme-mediated inactivation of mutant K-ras oncogene in a colon cancer cell line. Export

@article{citeulike:851142, abstract = {Mutation of c-K-ras oncogene is an important step in progression of colon cancer. We used a hammerhead ribozyme (KrasRz) against mutated K-ras gene transcripts (codon 12, GTT) to inactivate mutant K-ras function in the colon cancer cell line SW480, harbouring a mutant K-ras gene. The beta-actin promoter-driven KrasRz sequence (pHbeta/KrasRz) was introduced into these cells (SW480/KrasRz), and we evaluated its effects on growth of the colon cancer. The gene expression of angiogenesis-related molecules (vascular endothelial growth factor and thrombospondin) was also estimated in SW480/KrasRz. KrasRz specifically and efficiently cleaved the mutant K-ras mRNA but not wild-type mRNA in vitro. SW480/KrasRz showed decreased growth rate under tissue culture conditions (P< 0.01, Dunnett's test). The xenotransplantability of SW480/KrasRz (XeSW480/KrasRz) was significantly decreased in nude mice (P< 0.05, Fisher's exact test). Tumour volume of the xenografts XeSW480/KrasRz was significantly smaller than that of XeSW480/DisKrasRz (P< 0.01, Dunnett's test). Gene expression of VEGF was suppressed in SW480/KrasRz, while TSP1 gene expression was enhanced. The SW480/KrasRz cells showed apoptosis-related features including nuclear condensation and DNA fragmentation. These results suggested that the hammerhead ribozyme-mediated inactivation of the mutated K-ras mRNA induced growth suppression, apoptosis and alteration of angiogenic factor expression.}, address = {Department of Pathology, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa, 259-1193, Japan.}, author = {Tokunaga, T. and Tsuchida, T. and Kijima, H. and Okamoto, K. and Oshika, Y. and Sawa, N. and Ohnishi, Y. and Yamazaki, H. and Miura, S. and Ueyama, Y. and Nakamura, M.}, citeulike-article-id = {851142}, citeulike-linkout-0 = {http://dx.doi.org/10.1054/bjoc.2000.1363}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/10952790}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=10952790}, doi = {10.1054/bjoc.2000.1363}, issn = {0007-0920}, journal = {Br J Cancer}, keywords = {antisense, cancer, colon, gowth, inhibition, k-ras, mutation}, month = {September}, number = {6}, pages = {833--839}, posted-at = {2006-09-20 12:10:50}, priority = {2}, title = {Ribozyme-mediated inactivation of mutant K-ras oncogene in a colon cancer cell line.}, url = {http://dx.doi.org/10.1054/bjoc.2000.1363}, volume = {83}, year = {2000} }

TY - JOUR ID - citeulike:851142 L3 - citeulike-article-id:851142 N2 - Mutation of c-K-ras oncogene is an important step in progression of colon cancer. We used a hammerhead ribozyme (KrasRz) against mutated K-ras gene transcripts (codon 12, GTT) to inactivate mutant K-ras function in the colon cancer cell line SW480, harbouring a mutant K-ras gene. The beta-actin promoter-driven KrasRz sequence (pHbeta/KrasRz) was introduced into these cells (SW480/KrasRz), and we evaluated its effects on growth of the colon cancer. The gene expression of angiogenesis-related molecules (vascular endothelial growth factor and thrombospondin) was also estimated in SW480/KrasRz. KrasRz specifically and efficiently cleaved the mutant K-ras mRNA but not wild-type mRNA in vitro. SW480/KrasRz showed decreased growth rate under tissue culture conditions (P< 0.01, Dunnett's test). The xenotransplantability of SW480/KrasRz (XeSW480/KrasRz) was significantly decreased in nude mice (P< 0.05, Fisher's exact test). Tumour volume of the xenografts XeSW480/KrasRz was significantly smaller than that of XeSW480/DisKrasRz (P< 0.01, Dunnett's test). Gene expression of VEGF was suppressed in SW480/KrasRz, while TSP1 gene expression was enhanced. The SW480/KrasRz cells showed apoptosis-related features including nuclear condensation and DNA fragmentation. These results suggested that the hammerhead ribozyme-mediated inactivation of the mutated K-ras mRNA induced growth suppression, apoptosis and alteration of angiogenic factor expression. IS - 6 JF - Br J Cancer SN - 0007-0920 AD - Department of Pathology, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa, 259-1193, Japan. EP - 839 TI - Ribozyme-mediated inactivation of mutant K-ras oncogene in a colon cancer cell line. VL - 83 SP - 833 KW - antisense KW - cancer KW - colon KW - gowth KW - inhibition KW - k-ras KW - mutation AU - Tokunaga, T AU - Tsuchida, T AU - Kijima, H AU - Okamoto, K AU - Oshika, Y AU - Sawa, N AU - Ohnishi, Y AU - Yamazaki, H AU - Miura, S AU - Ueyama, Y AU - Nakamura, M PY - 2000/09// UR - http://dx.doi.org/10.1054/bjoc.2000.1363 ER -