E47 Governs the MYC-CDKN1B/p27KIP1-RB Network to Growth Arrest PDA Cells Independent of CDKN2A/p16INK4A and Wild-Type p53Summary
| Autor: | Pamela Itkin-Ansari, Roman Sasik, Yifan Wu, Reyhaneh Lahmy, Rachel Medal, Randall French, Kathleen M. Scully, Lia Signaevskaia, Brian P. James, Heejung Kim, Andrew M. Lowy |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Gene knockdown Small interfering RNA Hepatology Gastroenterology RNA Cell cycle Biology Cell morphology 3. Good health Cell biology Transcriptome Small hairpin RNA 03 medical and health sciences 030104 developmental biology lcsh:Diseases of the digestive system. Gastroenterology lcsh:RC799-869 Transcription factor |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology, Vol 6, Iss 2, Pp 181-198 (2018) |
| Popis: | Background & Aims: Oncogenic mutations in KRAS, coupled with inactivation of p53, CDKN2A/p16INK4A, and SMAD4, drive progression of pancreatic ductal adenocarcinoma (PDA). Overexpression of MYC and deregulation of retinoblastoma (RB) further promote cell proliferation and make identifying a means to therapeutically alter cell-cycle control pathways in PDA a significant challenge. We previously showed that the basic helix-loop-helix transcription factor E47 induced stable growth arrest in PDA cells in vitro and in vivo. Here, we identified molecular mechanisms that underlie E47-induced growth arrest in low-passage, patient-derived primary and established PDA cell lines. Methods: RNA sequencing was used to profile E47-dependent transcriptomes in 5 PDA cell lines. Gene Ontology analysis identified cell-cycle control as the most altered pathway. Small interfering RNA/short hairpin RNA knockdown, small-molecule inhibitors, and viral expression were used to examine the function of E47-dependent genes in cell-cycle arrest. Cell morphology, expression of molecular markers, and senescence-associated β-galactosidase activity assays identified cellular senescence. Results: E47 uniformly inhibited PDA cell-cycle progression by decreasing expression of MYC, increasing the level of CDKN1B/p27KIP1, and restoring RB tumor-suppressor function. The molecular mechanisms by which E47 elicited these changes included altering both RNA transcript levels and protein stability of MYC and CDKN1B/p27KIP1. At the cellular level, E47 elicited a senescence-like phenotype characterized by increased senescence-associated β-galactosidase activity and altered expression of senescence markers. Conclusions: E47 governs a highly conserved network of cell-cycle control genes, including MYC, CDKN1B/p27KIP1, and RB, which can induce a senescence-like program in PDA cells that lack CDKN2A/p16INK4A and wild-type p53. RNA sequencing data are available at the National Center for Biotechnology Information GEO at https://www.ncbi.nlm.nih.gov/geo/; accession number: GSE100327. Keywords: Pancreatic Ductal Adenocarcinoma, bHLH, Cell Cycle, Senescence |
| Databáze: | OpenAIRE |
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