title: Loss of cyclin g1 expression in human uterine leiomyoma cells induces apoptosis.
abstract: Observations from the authors' laboratory suggest a physiological role for increased cyclin G1 protein levels in human uterine leiomyoma. The hypothesis of the present study is that the strategic modulation of cyclin G1 by antisense technology will inhibit the survival of in vitro-grown uterine leiomyoma cells. Cultured uterine leiomyoma cells were transfected with cyclin G1 ribbon-type antisense oligonucleotide (cyclin G1 RiAS) to effectively reduce cyclin G1 expression. Cell viability, in situ terminal deoxyuridine nick end-labeling (TUNEL) assay, flow cytometry, DNA fragmentation, and expression of cell cycle regulatory-related proteins were evaluated by Western blot. Antisense oligonucleotides compromised uterine leiomyoma cell viability and inducted apoptosis in a caspase-independent mechanism. In situ TUNEL and DNA fragmentation revealed apoptosis induction, and fluorescent-activated cell sorting analysis showed increased sub-G1-phase cells. Furthermore, abrogation of cyclin G1 enhanced p53 accumulation, phosphorylation of p53 at Ser-15 residue, and increased expression of cyclin-dependent kinase inhibitors p21 and p27. These data imply that cyclin G1 expression is associated with growth promotion and the potential utility and novelty of using ribbon-type antisense oligonucleotides as a gene therapy strategy to treat human uterine leiomyoma.
authors.affiliation: Department of Obstetrics and Gynecology, School of Medicine, Keimyung University, Daegu, Korea.
authors.forename: Sang-Hoon
authors.initials: SH
authors.lastname: Kwon
authors.forename: Joon-Cheol
authors.initials: JC
authors.lastname: Park
authors.forename: Sabarish
authors.initials: S
authors.lastname: Ramachandran
authors.forename: Soon-Do
authors.initials: SD
authors.lastname: Cha
authors.forename: Kun-Young
authors.initials: KY
authors.lastname: Kwon
authors.forename: Jong-Ku
authors.initials: JK
authors.lastname: Park
authors.forename: Jong-Wook
authors.initials: JW
authors.lastname: Park
authors.forename: Insoo
authors.initials: I
authors.lastname: Bae
authors.forename: Chi-Heum
authors.initials: CH
authors.lastname: Cho
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D015153:Blotting, Western; D053148:Caspase 3 / Q000502:physiology; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D056746:Cyclin G; D056766:Cyclin G1; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D016213:Cyclins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D053938:DNA Fragmentation; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D020287:In Situ Nick-End Labeling; D007889:Leiomyoma / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D009215:Myometrium / Q000166:cytology / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014594:Uterine Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000628:therapy
chemical_list: C534702:CCNG1 protein, human; C534704:Ccng1 protein, rat; D018797:Cell Cycle Proteins; D056746:Cyclin G; D056766:Cyclin G1; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D016376:Oligonucleotides, Antisense; D016159:Tumor Suppressor Protein p53; D050760:Cyclin-Dependent Kinase Inhibitor p27; D053148:Caspase 3