title: Genomic instability and proliferation/survival pathways in RB1-deficient malignancies.
abstract: Genomic instability (GIN) is a hallmark of most cancer cells. However, compared to most human cancer cell types, the retinoblastoma tumor cells show a relatively stable genome. The fundamental basis of this genomic stability has yet to be elucidated, and the role of certain proteins involved in cell cycle regulation may be the key to the development of these specific genotypes. We examined whether thyroid hormone receptor beta 1 and 2 (TRβ1 and TRβ2), known to regulate tumorigenesis, and PTTG1, a mitotic checkpoint protein, play a role in maintaining genomic stability in retinoblastoma. In order to elucidate the role of these proteins in development of aneuploidy/polyploidy, an indicator of GIN, we first studied comparative GIN in retinoblastomas and multiple RB mutant cancer cell lines using single nucleotide polymorphism (SNP) analysis. We then utilized pLKO lentiviral vectors to selectively modify expression of the targeted cell cycle proteins and interpret their effect on downstream cell cycle proteins and their relative effects on the development of polyploidy in multiple tumor cell lines. The SNP analysis showed that retinoblastomas displayed relatively fewer genomic copy number changes as compared to other RB1-deficient cancer cell lines. Both TRβ1 and TRβ2 knockdown led to accumulation of E2F1 and PTTG1 and increased GIN as demonstrated by an increase in polyploidy. Downregulation of PTTG1 led to a relative decrease in GIN while upregulation of PTTG1 led to a relative increase in GIN. Knockdown of E2F1 led to a downstream decrease in PTTG1 expression. Rb-knockdown also upregulated E2F1 and PTTG1 leading to increased GIN. We showed that Rb is necessary for PTTG1 inhibition and genomic stability. A relatively stable genome in retinoblastoma tumor cells is maintained by TRβ1 and TRβ2-mediated PTTG1 inhibition, counteracting Rb-deficiency-related GIN. TRβ1, TRβ2 and Rb-KD all led to the downstream PTTG1 accumulation, apparently through an activation of E2F1 resulting in extensive genomic instability as seen in other Rb-deficient tumors.
authors.affiliation: Departments of Pathology, Pediatrics, Surgery, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, USA.
authors.forename: Lara
authors.initials: L
authors.lastname: Pappas
authors.affiliation: Departments of Pathology, Pediatrics, Surgery, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, USA.
authors.forename: Xiaoliang Leon
authors.initials: XL
authors.lastname: Xu
authors.affiliation: Departments of Pathology, Pediatrics, Surgery, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: Abramson
authors.affiliation: Departments of Pathology, Pediatrics, Surgery, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, USA. Electronic address: Jhanwars@mskcc.org.
authors.forename: Suresh C
authors.initials: SC
authors.lastname: Jhanwar
mesh_terms: D000818:Animals; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D050687:E2F1 Transcription Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D042822:Genomic Instability; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D046228:Microarray Analysis; D020641:Polymorphism, Single Nucleotide; D011123:Polyploidy; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D019572:Retinal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D012175:Retinoblastoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016160:Retinoblastoma Protein / Q000172:deficiency / Q000235:genetics*; D064249:Securin / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D037042:Thyroid Hormone Receptors beta / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; C578005:PTTG1 protein, mouse; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D016160:Retinoblastoma Protein; D064249:Securin; D037042:Thyroid Hormone Receptors beta
keywords: Genomic instability; Proliferation; Rb-deficient cancers; Retinoblastoma; Survival pathways