Hammerhead ribozyme-mediated inactivation of mutant RET in medullary thyroid carcinoma.
Activating mutations of the RET proto-oncogene cause hereditary medullary thyroid carcinoma. To examine whether selective inactivation of mutant RET could prevent transformation, a hammerhead ribozyme was designed to cleave RET mRNA containing a transforming mutation of codon 634 TGC --> TAC (Cys634Tyr). In vitro RNA cleavage assay demonstrated that the ribozyme selectively cleaved RET RNA with a Cys634Tyr but not Cys634Arg or the normal sequence. Expression of ribozyme in NIH/3T3 cells prevented RET-mediated colony formation in soft agar. This inhibition required catalytically active ribozyme and was specific for the TAC mutation. Therefore, ribozymes designed to selectively target mutant RET RNA may provide an effective therapeutic in the treatment of this syndrome.
Parthasarathy
R
R
Department of Internal Medicine Specialties, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Cote
G J
GJ
Gagel
R F
RF
D016475:3T3 Cells; D000818:Animals; D018276:Carcinoma, Medullary / Q000235:genetics*; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D029721:Drosophila Proteins; D015972:Gene Expression Regulation, Neoplastic; D051379:Mice; D009154:Mutation; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051096:Proto-Oncogene Proteins c-ret; D016337:RNA, Catalytic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D013964:Thyroid Neoplasms / Q000235:genetics*; D014407:Tumor Cells, Cultured
D029721:Drosophila Proteins; D011518:Proto-Oncogene Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila; C494999:Ret protein, mouse