Document Detail


Telomerase inhibition by an siRNA directed against hTERT leads to telomere attrition in HT29 cells.
MedLine Citation:
PMID:  16820926     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human telomerase is a ribonucleoprotein complex composed of at least the reverse catalytic transcriptase hTERT and RNA component hTR. The enzyme stabilizes telomere length and thereby contributes to unlimited cell proliferation, i.e. immortality. Reactivation of telomerase activity during carcinogenesis is a common hallmark in most human tumor types. Consequently, telomerase is an attractive molecular target toward which to direct cancer therapeutic agents. RNA interference (RNAi) has been shown to be an effective method for inhibiting the expression of a given gene in human cells by targeting with short duplex RNA (short-interfering RNA or siRNA). Thus, we evaluated the ability of siRNAs to inhibit telomerase activity in the HT29 immortal human colorectal adenocarcinoma cell line as a model for colorectal carcinogenesis. By transient expression of a specific siRNA directed against hTERT, we reduced telomerase activity in the transfected cells. Moreover, telomere lengths were reduced in cells stably expressing this particular RNA sequence, cloned as an shRNA into an eukaryotic expression vector. The cell clone that displayed a telomerase-negative phenotype showed dramatically reduced telomere lengths and stopped proliferation. We observed that the vector was integrated into the cell genome and, despite telomere shortening, cells retained their MSI phenotype. We conclude that we have developed a potent telomerase inhibitor leading to cell death.
Authors:
Patrícia de Souza Nascimento; Gilda Alves; Wolfgang Fiedler
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Oncology reports     Volume:  16     ISSN:  1021-335X     ISO Abbreviation:  Oncol. Rep.     Publication Date:  2006 Aug 
Date Detail:
Created Date:  2006-07-05     Completed Date:  2006-09-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9422756     Medline TA:  Oncol Rep     Country:  Greece    
Other Details:
Languages:  eng     Pagination:  423-8     Citation Subset:  IM    
Affiliation:
Klinik und Poliklinik f. Innere Medizin I - Sektion Molekulare Gastroenterologische Onkologie, Martin Luther Universität Halle-Wittenberg, Halle, Germany.
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MeSH Terms
Descriptor/Qualifier:
Adenocarcinoma / enzymology*,  genetics
Colorectal Neoplasms / enzymology*,  genetics
DNA-Binding Proteins / antagonists & inhibitors*,  genetics
Genomic Instability
Humans
Microsatellite Repeats
RNA Interference
RNA, Messenger / analysis,  metabolism
RNA, Small Interfering / genetics,  pharmacology*
Telomerase / antagonists & inhibitors*,  genetics
Telomere / metabolism*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/DNA-Binding Proteins; 0/RNA, Messenger; 0/RNA, Small Interfering; EC 2.7.7.49/Telomerase

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