Oncotarget

Research Papers:

Gene expression profiling of tumor-initiating stem cells from mouse Krebs-2 carcinoma using a novel marker of poorly differentiated cells

Ekaterina A. Potter, Evgenia V. Dolgova, Anastasia S. Proskurina, Yaroslav R. Efremov, Alexandra M. Minkevich, Aleksey S. Rozanov, Sergey E. Peltek, Valeriy P. Nikolin, Nelly A. Popova, Igor A. Seledtsov, Vladimir V. Molodtsov, Evgeniy L. Zavyalov, Oleg S. Taranov, Sergey I. Baiborodin, Alexander A. Ostanin, Elena R. Chernykh, Nikolay A. Kolchanov and Sergey S. Bogachev _

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Oncotarget. 2017; 8:9425-9441. https://doi.org/10.18632/oncotarget.14116

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Abstract

Ekaterina A. Potter1,*, Evgenia V. Dolgova1,*, Anastasia S. Proskurina1,*, Yaroslav R. Efremov1,2, Alexandra M. Minkevich1, Aleksey S. Rozanov1, Sergey E. Peltek1, Valeriy P. Nikolin1, Nelly A. Popova1,2, Igor A. Seledtsov3, Vladimir V. Molodtsov2,3, Evgeniy L Zavyalov1, Oleg S. Taranov4, Sergey I. Baiborodin1, Alexander A. Ostanin5, Elena R. Chernykh5, Nikolay A. Kolchanov1, Sergey S. Bogachev1

1Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia

2Novosibirsk State University, Novosibirsk 630090, Russia

3Softberry Inc., New York 10549, USA

4The State Research Center of Virology and Biotechnology VECTOR, Koltsovo, Novosibirsk 630559, Russia

5Research Institute of Fundamental and Clinical Immunology, Novosibirsk 630099, Russia

*These authors contributed equally to this work

Correspondence to:

Sergey S. Bogachev, email: labmolbiol@mail.ru

Keywords: tumor-initiating stem cells, DNA internalization, RNAseq, Real Time PCR, TAMRA

Received: August 29, 2016     Accepted: December 15, 2016     Published: December 23, 2016

ABSTRACT

Using the ability of poorly differentiated cells to natively internalize fragments of extracellular double-stranded DNA as a marker, we isolated a tumorigenic subpopulation present in Krebs-2 ascites that demonstrated the features of tumor-inducing cancer stem cells. Having combined TAMRA-labeled DNA probe and the power of RNA-seq technology, we identified a set of 168 genes specifically expressed in TAMRA-positive cells (tumor-initiating stem cells), these genes remaining silent in TAMRA-negative cancer cells. TAMRA+ cells displayed gene expression signatures characteristic of both stem cells and cancer cells. The observed expression differences between TAMRA+ and TAMRA− cells were validated by Real Time PCR. The results obtained corroborated the biological data that TAMRA+ murine Krebs-2 tumor cells are tumor-initiating stem cells. The approach developed can be applied to profile any poorly differentiated cell types that are capable of immanent internalization of double-stranded DNA.


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