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TKTL1 expression in human malign and benign cell lines

BACKGROUND: Overexpression of transketolase-like 1 protein TKTL1 in cancer cells has been reported to correlate with enhanced glycolysis and lactic acid production. Furthermore, enhanced TKTL1 expression was put into context with resistance to chemotherapy and ionizing radiation. Here, a panel of hu...

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Autores principales: Kämmerer, Ulrike, Gires, Olivier, Pfetzer, Nadja, Wiegering, Armin, Klement, Rainer Johannes, Otto, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4506423/
https://www.ncbi.nlm.nih.gov/pubmed/26187043
http://dx.doi.org/10.1186/1471-2407-15-2
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author Kämmerer, Ulrike
Gires, Olivier
Pfetzer, Nadja
Wiegering, Armin
Klement, Rainer Johannes
Otto, Christoph
author_facet Kämmerer, Ulrike
Gires, Olivier
Pfetzer, Nadja
Wiegering, Armin
Klement, Rainer Johannes
Otto, Christoph
author_sort Kämmerer, Ulrike
collection PubMed
description BACKGROUND: Overexpression of transketolase-like 1 protein TKTL1 in cancer cells has been reported to correlate with enhanced glycolysis and lactic acid production. Furthermore, enhanced TKTL1 expression was put into context with resistance to chemotherapy and ionizing radiation. Here, a panel of human malign and benign cells, which cover a broad range of chemotherapy and radiation resistance as well as reliance on glucose metabolism, was analyzed in vitro for TKTL1 expression. METHODS: 17 malign and three benign cell lines were characterized according to their expression of TKTL1 on the protein level with three commercially available anti-TKTL1 antibodies utilizing immunohistochemistry and Western blot, as well as on mRNA level with three published primer pairs for RT-qPCR. Furthermore, sensitivities to paclitaxel, cisplatin and ionizing radiation were assessed in cell survival assays. Glucose consumption and lactate production were quantified as surrogates for the “Warburg effect”. RESULTS: Considerable amounts of tktl1 mRNA and TKTL1 protein were detected only upon stable transfection of the human embryonic kidney cell line HEK293 with an expression plasmid for human TKTL1. Beyond that, weak expression of endogenous tktl1 mRNA was measured in the cell lines JAR and U251. Western blot analysis of JAR and U251 cells did not detect TKTL1 at the expected size of 65 kDa with all three antibodies specific for TKTL1 protein and immunohistochemical staining was observed with antibody JFC12T10 only. All other cell lines tested here revealed expression of tktl1 mRNA below detection limits and were negative for TKTL1 protein. However, in all cell lines including TKTL1-negative HEK293-control cells, antibody JFC12T10 detected multiple proteins with different molecular weights. Importantly, JAR and U251 did neither demonstrate an outstanding production of lactic acid nor increased resistance against chemotherapeutics or to ionizing radiation, respectively. CONCLUSION: Using RT-qPCR and three different antibodies we observed only exceptional occurrence of TKTL1 in a panel of malignant human cell lines in vitro. The presence of TKTL1 was unrelated to either the rate of glucose consumption/lactic acid production or resistance against chemo- and radiotherapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2407-15-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-45064232015-07-19 TKTL1 expression in human malign and benign cell lines Kämmerer, Ulrike Gires, Olivier Pfetzer, Nadja Wiegering, Armin Klement, Rainer Johannes Otto, Christoph BMC Cancer Research Article BACKGROUND: Overexpression of transketolase-like 1 protein TKTL1 in cancer cells has been reported to correlate with enhanced glycolysis and lactic acid production. Furthermore, enhanced TKTL1 expression was put into context with resistance to chemotherapy and ionizing radiation. Here, a panel of human malign and benign cells, which cover a broad range of chemotherapy and radiation resistance as well as reliance on glucose metabolism, was analyzed in vitro for TKTL1 expression. METHODS: 17 malign and three benign cell lines were characterized according to their expression of TKTL1 on the protein level with three commercially available anti-TKTL1 antibodies utilizing immunohistochemistry and Western blot, as well as on mRNA level with three published primer pairs for RT-qPCR. Furthermore, sensitivities to paclitaxel, cisplatin and ionizing radiation were assessed in cell survival assays. Glucose consumption and lactate production were quantified as surrogates for the “Warburg effect”. RESULTS: Considerable amounts of tktl1 mRNA and TKTL1 protein were detected only upon stable transfection of the human embryonic kidney cell line HEK293 with an expression plasmid for human TKTL1. Beyond that, weak expression of endogenous tktl1 mRNA was measured in the cell lines JAR and U251. Western blot analysis of JAR and U251 cells did not detect TKTL1 at the expected size of 65 kDa with all three antibodies specific for TKTL1 protein and immunohistochemical staining was observed with antibody JFC12T10 only. All other cell lines tested here revealed expression of tktl1 mRNA below detection limits and were negative for TKTL1 protein. However, in all cell lines including TKTL1-negative HEK293-control cells, antibody JFC12T10 detected multiple proteins with different molecular weights. Importantly, JAR and U251 did neither demonstrate an outstanding production of lactic acid nor increased resistance against chemotherapeutics or to ionizing radiation, respectively. CONCLUSION: Using RT-qPCR and three different antibodies we observed only exceptional occurrence of TKTL1 in a panel of malignant human cell lines in vitro. The presence of TKTL1 was unrelated to either the rate of glucose consumption/lactic acid production or resistance against chemo- and radiotherapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2407-15-2) contains supplementary material, which is available to authorized users. BioMed Central 2015-06-10 /pmc/articles/PMC4506423/ /pubmed/26187043 http://dx.doi.org/10.1186/1471-2407-15-2 Text en © Kämmerer et al. 2015 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Kämmerer, Ulrike
Gires, Olivier
Pfetzer, Nadja
Wiegering, Armin
Klement, Rainer Johannes
Otto, Christoph
TKTL1 expression in human malign and benign cell lines
title TKTL1 expression in human malign and benign cell lines
title_full TKTL1 expression in human malign and benign cell lines
title_fullStr TKTL1 expression in human malign and benign cell lines
title_full_unstemmed TKTL1 expression in human malign and benign cell lines
title_short TKTL1 expression in human malign and benign cell lines
title_sort tktl1 expression in human malign and benign cell lines
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4506423/
https://www.ncbi.nlm.nih.gov/pubmed/26187043
http://dx.doi.org/10.1186/1471-2407-15-2
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