Di Cecilia, Serena
(2015)
A Novel Long Non-Coding RNA Transcript Regulates The Fate of Colon Cancer Initiation.
[Tesi di dottorato]
| Item Type: |
Tesi di dottorato
|
| Resource language: |
English |
| Title: |
A Novel Long Non-Coding RNA Transcript Regulates The Fate of Colon Cancer Initiation |
| Creators: |
| Creators | Email |
|---|
| Di Cecilia, Serena | serenadicecilia@gmail.com |
|
| Date: |
2 March 2015 |
| Number of Pages: |
108 |
| Institution: |
Università degli Studi di Napoli Federico II |
| Department: |
Scienze Mediche Traslazionali |
| Scuola di dottorato: |
SEMM - European School of Molecular Medicine |
| Dottorato: |
PhD in Molecular Medicine (Molecular Oncology or Human Genetics) |
| Ciclo di dottorato: |
26 |
| Coordinatore del Corso di dottorato: |
| nome | email |
|---|
| Ballabio, Andrea | ballabio@igem.it |
|
| Tutor: |
| nome | email |
|---|
| Salvatore, Francesco | UNSPECIFIED | | Del Vecchio, Luigi | UNSPECIFIED | | Martin, Walsh | UNSPECIFIED |
|
| Date: |
2 March 2015 |
| Number of Pages: |
108 |
| Keywords: |
lncRNA, Colon Cancer initiating cells, Wnt-signalling, LUST |
| Settori scientifico-disciplinari del MIUR: |
Area 05 - Scienze biologiche > BIO/12 - Biochimica clinica e biologia molecolare clinica
Area 06 - Scienze mediche > MED/03 - Genetica medica |
[error in script]
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| Date Deposited: |
12 Jun 2015 10:11 |
| Last Modified: |
31 Dec 2016 02:00 |
| URI: |
http://www.fedoa.unina.it/id/eprint/10058 |
| DOI: |
10.6092/UNINA/FEDOA/10058 |
Collection description
Cancer Initiating cells are a small population of cancer cells capable of tumor initiation and growth. Characteristic of these cells are the expression of “stem-like” markers and the common alteration of the Wnt/β-catenin and Notch pathways. In my thesis, I characterized Colon Cancer Initiating Cells (CCICs) by FACS analysis from HT-29 colon cancer cell line. I then performed in vitro colonospheres-forming assay and RNA-Seq, to interrogate the genome-wide signature involving CCICs. My results show that members of Wnt/β-catenin pathway are elevated in CCICs and colonospheres, when compared to a more differentiated population. Moreover, genes directing the differentiation were silenced in CCICs and colonospheres. These results demonstrate the self-renewal and proliferation capacity of the isolated CCICs population, and that this subpopulation resembles a strong Wnt-signaling pathway signature. Additionally, several lncRNAs are dys-regulated in CCICs, and my findings identify LUST/RBM5-AS1 as a lncRNA transcript strongly elevated during colonospheres formation. The expression of LUST and stem-like markers CD24 and CD44, and expression of Wnt-signaling corresponds with cells that can survive and grow in serum-free media. Loss of LUST impairs Wnt-signaling at mRNA and protein levels, while LUST overexpression provides enhanced and synergistic signaling mediated through Wnt and β-catenin at the mRNA and protein levels. Nuclear/Cytoplasmic RNA fractionation and RNA-FISH show that LUST essentially is a nuclear transcript. RNA immuno-precipitation and UV cross-linking immuno-precipitation assays show that LUST RNA binds to β-catenin. Finally, LUST overexpression enhances colonospheres formation more rapidly. Collectively, my findings reveal that the lncRNA LUST regulates Wnt pathway in CCICs through a coordinated physical interaction with β-catenin, acting through transcriptional regulation to promote colon cancer initiating cells maintenance.
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