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| Status |
Public on Jun 09, 2010 |
| Title |
Comparison of CD44+/CD24- sorted fraction with 'rest' fraction of NBLE cells and primary breast cancer-derived cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
TSV40ER and hTERT immortalized mammospheres undergo transformation in culture to generate breast cancer stem cells
The stem cell hypothesis of cancer predicts that cancers arise from the transformation of normal stem/progenitor cells present within adult tissues. However, the molecular mechanisms that lead to the transformation of these cells remain largely unknown, mainly owing to the lack of suitable model systems. In this study, we have generated an immortalized breast epithelial cell line, NBLE, by over-expressing the SV40 Early Region and the catalytic domain of human telomerase into mammosphere-derived cells that are enriched in breast stem/progenitor cells. While primary mammospheres fail to grow beyond four passages in suspension culture, NBLE cells propagated continuously generating spheres at a high frequency of 35%. These cells also expressed stemness-related genes, and differentiated into both luminal and myoepithelial lineages. Interestingly, after around 135 population doublings in culture, the late passage cells exhibited a mesenchymal morphology and contained >90% of CD44+/CD24Low/- cells resembling breast cancer stem cells. When injected subcutaneously into nude mice, the late passage NBLEs formed invasive tumors closely resembling breast adenocarcinomas, the most common type of breast cancer. The transformed NBLE cells showed hyperactivation of Wnt, Hh and TGFbeta pathways, suggesting that the deregulation of these self-renewal pathways may have in part contributed to tumorigenesis. Gene expression analysis further revealed a high degree of similarity with breast cancer stem cells. Thus, our study provides a unique model system to study signaling pathways involved in self-renewal, differentiation, and transformation of breast stem/progenitor cells. Additionally, the NBLE cells can be utilized for screening drugs specifically targeting breast cancer stem cells.
NBLE cells were generated by over-expressing SV40ER and hTERT into mammosphere-derived cells. These cells harnored CD44+/CD24- fraction that resembled breast cancer stem cells. To understand the unique gene expression of this fraction we performed microarray on CD44+/CD24- versus rest fractions from NBLE cells (Rep 1 and Rep 2). To see if this gene profile has any similarity with breast cancer stem cells, we performed CD44+/CD24- versus rest microarray on two primary breast tumors (CB272 and CB258-Rep1 andRep2).
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| Overall design |
Whole genome gene expression profiling microarray based comparison of immortalized and transformed mammospheres derived cells.
Two-color experiment, NBLE-CD44+/CD24- labelled with Cy5 and NBLE-Rest labelled with Cy3
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| Contributor(s) |
Paranjape AN, Mandal T, Mukherjee G, Rangarajan A |
| Citation missing |
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| Submission date |
Jun 07, 2010 |
| Last update date |
Jan 23, 2019 |
| Contact name |
Anurag N Paranjape |
| E-mail(s) |
anurag@mrdg.iisc.ernet.in
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| Phone |
+9180-22933263
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| Organization name |
Indian Institute of Science
|
| Department |
MRDG
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| Lab |
Dr Annapoorni Rangarajan's lab
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| Street address |
CVRaman road
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| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560012 |
| Country |
India |
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| Platforms (2) |
| GPL6480 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Probe Name version) |
| GPL10487 |
Genotypic Designed Human Whole genome gene expression profiling microarray 4x44k |
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| Samples (7)
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| GSM551969 |
Immortalized Human Mammary Epithelial Cells - Early Passage |
| GSM551970 |
Transformed Human Mammary Epithelial Cells - Late Passage |
| GSM622336 |
NBLE-CD44+/CD24- Rep 1 |
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| Relations |
| BioProject |
PRJNA127687 |
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