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Reprogramming bladder cancer cells for studying cancer initiation and progression

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Tumor Biology

Abstract

The induced pluripotent stem cell (iPSC) technology is the forced expression of specific transcription factors in somatic cells resulting in transformation into self-renewing, pluripotent cells which possess the ability to differentiate into any type of cells in the human body. While malignant cells could also be reprogrammed into iPSC-like cells with lower efficiency due to the genetic and epigenetic barriers in cancer cells, only a limited number of cancer cell types could be successfully reprogrammed until today. In the present study, we aimed at reprogramming two bladder cancer cell lines HTB-9 and T24 using a non-integrating Sendai virus (SeV) system. We have generated six sub-clones using distinct combinations of four factors—OCT4, SOX2, KLF4 and c-MYC—in two bladder cancer cell lines. Only a single sub-clone, T24 transduced with 4Fs, gave rise to iPSC-like cells. Bladder cancer cell-derived T24 4F cells represent unique features of pluripotent cells such as epithelial-like morphology, colony-forming ability, expression of pluripotency-associated markers and bearing the ability to differentiate in vitro. This is the first study focusing on the reprogramming susceptibility of two different bladder cancer cell lines to nuclear reprogramming. Further molecular characterisation of T24 4F cells could provide a better insight for biomarker research in bladder carcinogenesis and could offer a valuable tool for the development of novel therapeutic approaches in bladder carcinoma.

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Authors and Affiliations

  1. Department of Medical Biology, Faculty of Medicine, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Banu Iskender & Halit Canatan

  2. Betul-Ziya Eren Genome and Stem Cell Centre, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Banu Iskender, Kenan Izgi & Halit Canatan

  3. Department of Medical Biochemistry, Faculty of Medicine, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Kenan Izgi

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  1. Banu Iskender
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Correspondence to Banu Iskender.

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This study was supported by the grants from The Scientific and Technological Research Council of Turkey (No: 114S542 and 113S927).

Electronic supplementary material

Supplementary Figure 1

Immunofluorescent analysis of pluripotency-associated markers NANOG, OCT4, SOX2, TRA-1-60 and SSEA-4 in untransduced parental T24 and T24 cells transduced with 4Fs. Transduced T24 4F cells organised as colonies which were positively stained for pluripotency-associated markers while parental T24 cells exhibited a restricted expression in monolayer cultures. Nuclei were stained with DAPI. Scale bar represents 100 μm. (PDF 18941 kb) (PDF 18941 kb)

Supplementary Figure 2

Immunofluorescent analysis of pluripotency-associated markers NANOG, OCT4, SOX2, SSEA-4 and TRA-1-60 in untransduced parental HTB-9 cells and HTB-9 cells transduced with 4Fs. Untransduced parental T24 cells were compared to transduced HTB-9 4F cells at passage 10. Although transduction of HTB-9 cells resulted in an increase in expressions of pluripotency-associated markers SOX2, TRA-1-60 and SSEA-4, cells did not form colonies or represent typical colony formation of pluripotent cells. Nuclei were stained with DAPI. Scale bar represents 100 μm. (PDF 17820 kb)

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Iskender, B., Izgi, K. & Canatan, H. Reprogramming bladder cancer cells for studying cancer initiation and progression. Tumor Biol. 37, 13237–13245 (2016). https://doi.org/10.1007/s13277-016-5226-4

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