title: Small antisense RNA to cyclin D1 generated by pre-tRNA splicing inhibits growth of human hepatoma cells.
abstract: Introns are present in some human pre-tRNAs. They are spliced out during the maturation processes of pre-tRNAs in a way that is irrelevant to their specific nucleotide sequences. This unique characteristic of tRNA splicing can be used for generation of small antisense RNAs by replacing the intron sequences with corresponding antisense sequences. In this work, the intron sequence of human pre-tRNAtyr gene was replaced with a 20 bp antisense sequence targeted to the 5' coding region of cyclin D1, a molecule that was over-expressed in many malignant proliferating cells. Under the control of U6 SnRNA promoter to further enhance transcription efficiency of the modified pre-tRNAtyr gene and subsequent antisense generation, the antisense RNA exhibited obvious suppression of cyclin D1 expression in H22 hepatoma cells. The growth of H22-transplanted tumors in mice was significantly inhibited when treated with naked plasmid DNA harboring the cyclin D1 antisense RNA generating cassette. Such tumor growth inhibition might be due to apoptosis caused by reduced cyclin D1 expression as revealed by immunohistochemical analysis of tumor samples.
authors.affiliation: Beijing Institute of Microbiology and Epidemiology, 20 Dongdajie, Fengtai, Beijing 100071, China.
authors.forename: Dazhi
authors.initials: D
authors.lastname: Lai
authors.forename: Shaojie
authors.initials: S
authors.lastname: Weng
authors.forename: Cui'e
authors.initials: C
authors.lastname: Wang
authors.forename: Lianquan
authors.initials: L
authors.lastname: Qi
authors.forename: Changming
authors.initials: C
authors.lastname: Yu
authors.forename: Ling
authors.initials: L
authors.lastname: Fu
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D002455:Cell Division; D045744:Cell Line, Tumor; D019938:Cyclin D1 / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics*; D016372:RNA, Antisense / Q000235:genetics*; D012365:RNA, Transfer, Tyr / Q000235:genetics; D014158:Transcription, Genetic; D014183:Transplantation, Heterologous
chemical_list: D004275:DNA, Ribosomal; D009838:Oligodeoxyribonucleotides; D012322:RNA Precursors; D016372:RNA, Antisense; D012365:RNA, Transfer, Tyr; D019938:Cyclin D1