Directly to content
  1. Publishing |
  2. Search |
  3. Browse |
  4. Recent items rss |
  5. Open Access |
  6. Jur. Issues |
  7. DeutschClear Cookie - decide language by browser settings

A mammosphere formation RNAi screen reveals that ATG4A promotes a breast cancer stem-like phenotype

Wolf, Jonas ; Dewi, Dyah Laksmi ; Fredebohm, Johannes ; Müller-Decker, Karin ; Flechtenmacher, Christa ; Hoheisel, Jörg D. ; Böttcher, Michael

In: Breast Cancer Research, 15 (2013), Nr. R109. pp. 1-13. ISSN 1465-542X

[img]
Preview
PDF, English
Download (2MB) | Lizenz: Creative Commons LizenzvertragA mammosphere formation RNAi screen reveals that ATG4A promotes a breast cancer stem-like phenotype by Wolf, Jonas ; Dewi, Dyah Laksmi ; Fredebohm, Johannes ; Müller-Decker, Karin ; Flechtenmacher, Christa ; Hoheisel, Jörg D. ; Böttcher, Michael underlies the terms of Creative Commons Attribution 3.0 Germany

Citation of documents: Please do not cite the URL that is displayed in your browser location input, instead use the DOI, URN or the persistent URL below, as we can guarantee their long-time accessibility.

Abstract

Introduction: Breast cancer stem cells are suspected to be responsible for tumour recurrence, metastasis formation as well as chemoresistance. Consequently, great efforts have been made to understand the molecular mechanisms underlying cancer stem cell maintenance. In order to study these rare cells in-vitro, they are typically enriched via mammosphere culture. Here we developed a mammosphere-based negative selection shRNAi screening system suitable to analyse the involvement of thousands of genes in the survival of cells with cancer stem cell properties. Methods: We describe a sub-population expressing the stem-like marker CD44+/CD24-/low in SUM149 that were enriched in mammospheres. To identify genes functionally involved in the maintenance of the sub-population with cancer stem cell properties, we targeted over 5000 genes by RNAi and tested their ability to grow as mammospheres. The identified candidate ATG4A was validated in mammosphere and soft agar colony formation assays. Further, we evaluated the influence of ATG4A expression on the sub-population expressing the stem-like marker CD44+/CD24low. Next, the tumorigenic potential of SUM149 after up- or down-regulation of ATG4A was examined by xenograft experiments. Results: Using this method, Jak-STAT as well as cytokine signalling were identified to be involved in mammosphere formation. Furthermore, the autophagy regulator ATG4A was found to be essential for the maintenance of a sub-population with cancer stem cell properties and to regulate breast cancer cell tumourigenicity in vivo. Conclusion: In summary, we present a high-throughput screening system to identify genes involved in cancer stem cell maintenance and demonstrate its utility by means of ATG4A.

Item Type: Article
Journal or Publication Title: Breast Cancer Research
Volume: 15
Number: R109
Publisher: BioMed Central
Place of Publication: London
Date Deposited: 27 Jan 2017 09:45
Date: 2013
ISSN: 1465-542X
Page Range: pp. 1-13
Faculties / Institutes: Service facilities > German Cancer Research Center (DKFZ)
Medizinische Fakultät Heidelberg > Pathologisches Institut
Subjects: 610 Medical sciences Medicine
About | FAQ | Contact | Imprint |
OA-LogoDINI certificate 2013Logo der Open-Archives-Initiative