TKTL1 expression in human malign and benign cell lines

BMC Cancer

Research Article

Armin Wiegering¹  Ulrike Kämmerer²  Nadja Pfetzer³  Olivier Gires⁴  Rainer Johannes Klement⁵  Christoph Otto⁶ 

[1] Department of General, Visceral, Vascular and Paediatric Surgery, University Hospital of Würzburg, Oberdürrbacher Str. 6, D-97080, Würzburg, Germany;Department of Obstetrics and Gynaecology, University of Würzburg Hospital, Josef-Schneider-Str. 4, D-97080, Würzburg, Germany;Department of Obstetrics and Gynaecology, University of Würzburg Hospital, Josef-Schneider-Str. 4, D-97080, Würzburg, Germany;Apoptosis and Tumour Metabolism Lab, CRUK Beatson Institute, Garscube Estate, Switchback Road, G61 1BD, Glasgow, United Kingdom;Department of Otorhinolaryngology, Head and Neck Surgery, Ludwig-Maximilians-University, Marchioninistr. 15, D-81377, Munich, Germany;Department of Radiotherapy and Radiation Oncology, Leopoldina Hospital Schweinfurt, Gustav-Adolf-Straße 8, D-97422, Schweinfurt, Germany;Experimental Surgery, Experimental Transplantation Immunology, Department of General, Visceral, Vascular and Paediatric Surgery, University Hospital of Würzburg, Oberdürrbacher Str. 6, D-97080, Würzburg, Germany;

关键词: Cancer cell lines;    TKTL1;    Immunohistochemistry;    RT-qPCT;   

DOI  :  10.1186/1471-2407-15-2

 received in 2014-06-12, accepted in 2014-11-27,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundOverexpression 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.Methods17 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”.ResultsConsiderable 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.ConclusionUsing 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.

【 授权许可】

CC BY   
© Kämmerer et al. 2015