Telomere Transcripts Target Telomerase in Human Cancer Cells
Long non-coding transcripts from telomeres, called telomeric repeat-containing RNA (TERRA), were identified as blocking telomerase activity (TA), a telomere maintenance mechanism (TMM), in tumors. We expressed recombinant TERRA transcripts in tumor cell lines with TA and with alternative lengthening...
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doaj-8decb998779d44ccbc1e6efed6dad4602020-11-24T21:24:55ZengMDPI AGGenes2073-44252016-08-01784610.3390/genes7080046genes7080046Telomere Transcripts Target Telomerase in Human Cancer CellsTheresa Kreilmeier0Doris Mejri1Marlene Hauck2Miriam Kleiter3Klaus Holzmann4Institute of Cancer Research, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, Vienna 1090, AustriaInstitute of Cancer Research, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, Vienna 1090, AustriaDepartment of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USADepartment for Companion Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, Vienna 1210, AustriaInstitute of Cancer Research, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, Vienna 1090, AustriaLong non-coding transcripts from telomeres, called telomeric repeat-containing RNA (TERRA), were identified as blocking telomerase activity (TA), a telomere maintenance mechanism (TMM), in tumors. We expressed recombinant TERRA transcripts in tumor cell lines with TA and with alternative lengthening of telomeres (ALT) to study effects on TMM and cell growth. Adeno- and lentivirus constructs (AV and LV) were established for transient and stable expression of approximately 130 units of telomere hexanucleotide repeats under control of cytomegalovirus (CMV) and human RNase P RNA H1 (hH1) promoters with and without polyadenylation, respectively. Six human tumor cell lines either using telomerase or ALT were infected and analyzed for TA levels. Pre-infection cells using telomerase had 1%–3% of the TERRA expression levels of ALT cells. AV and LV expression of recombinant TERRA in telomerase positive cells showed a 1.3–2.6 fold increase in TERRA levels, and a decrease in TA of 25%–58%. Dominant-negative or small hairpin RNA (shRNA) viral expression against human telomerase reverse transcriptase (hTERT) results in senescence, not induced by TERRA expression. Population doubling time, cell viability and TL (telomere length) were not impacted by ectopic TERRA expression. Clonal growth was reduced by TERRA expression in TA but not ALT cell lines. ALT cells were not affected by treatments applied. Established cell models and tools may be used to better understand the role of TERRA in the cell, especially for targeting telomerase.http://www.mdpi.com/2073-4425/7/8/46telomeraseenzyme inhibitiontelomerelong non-coding transcriptviral expression systemstumor cell lineshumancanine |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Theresa Kreilmeier Doris Mejri Marlene Hauck Miriam Kleiter Klaus Holzmann |
spellingShingle |
Theresa Kreilmeier Doris Mejri Marlene Hauck Miriam Kleiter Klaus Holzmann Telomere Transcripts Target Telomerase in Human Cancer Cells Genes telomerase enzyme inhibition telomere long non-coding transcript viral expression systems tumor cell lines human canine |
author_facet |
Theresa Kreilmeier Doris Mejri Marlene Hauck Miriam Kleiter Klaus Holzmann |
author_sort |
Theresa Kreilmeier |
title |
Telomere Transcripts Target Telomerase in Human Cancer Cells |
title_short |
Telomere Transcripts Target Telomerase in Human Cancer Cells |
title_full |
Telomere Transcripts Target Telomerase in Human Cancer Cells |
title_fullStr |
Telomere Transcripts Target Telomerase in Human Cancer Cells |
title_full_unstemmed |
Telomere Transcripts Target Telomerase in Human Cancer Cells |
title_sort |
telomere transcripts target telomerase in human cancer cells |
publisher |
MDPI AG |
series |
Genes |
issn |
2073-4425 |
publishDate |
2016-08-01 |
description |
Long non-coding transcripts from telomeres, called telomeric repeat-containing RNA (TERRA), were identified as blocking telomerase activity (TA), a telomere maintenance mechanism (TMM), in tumors. We expressed recombinant TERRA transcripts in tumor cell lines with TA and with alternative lengthening of telomeres (ALT) to study effects on TMM and cell growth. Adeno- and lentivirus constructs (AV and LV) were established for transient and stable expression of approximately 130 units of telomere hexanucleotide repeats under control of cytomegalovirus (CMV) and human RNase P RNA H1 (hH1) promoters with and without polyadenylation, respectively. Six human tumor cell lines either using telomerase or ALT were infected and analyzed for TA levels. Pre-infection cells using telomerase had 1%–3% of the TERRA expression levels of ALT cells. AV and LV expression of recombinant TERRA in telomerase positive cells showed a 1.3–2.6 fold increase in TERRA levels, and a decrease in TA of 25%–58%. Dominant-negative or small hairpin RNA (shRNA) viral expression against human telomerase reverse transcriptase (hTERT) results in senescence, not induced by TERRA expression. Population doubling time, cell viability and TL (telomere length) were not impacted by ectopic TERRA expression. Clonal growth was reduced by TERRA expression in TA but not ALT cell lines. ALT cells were not affected by treatments applied. Established cell models and tools may be used to better understand the role of TERRA in the cell, especially for targeting telomerase. |
topic |
telomerase enzyme inhibition telomere long non-coding transcript viral expression systems tumor cell lines human canine |
url |
http://www.mdpi.com/2073-4425/7/8/46 |
work_keys_str_mv |
AT theresakreilmeier telomeretranscriptstargettelomeraseinhumancancercells AT dorismejri telomeretranscriptstargettelomeraseinhumancancercells AT marlenehauck telomeretranscriptstargettelomeraseinhumancancercells AT miriamkleiter telomeretranscriptstargettelomeraseinhumancancercells AT klausholzmann telomeretranscriptstargettelomeraseinhumancancercells |
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