Highly sensitive profiling of CD44+/CD24- breast cancer stem cells by combining global mRNA amplification and next generation sequencing: evidence for a hyperactive PI3K pathway

Cancer Lett. 2012 Dec 28;325(2):165-74. doi: 10.1016/j.canlet.2012.06.010. Epub 2012 Jul 4.

Abstract

We performed next generation sequencing- and microarray-based gene expression profiling of CD44(+)/CD24(-)/CD45(-) breast CSCs (cancer stem cells) isolated from primary ERα-positive breast cancer. By combining semi-automated dissociation of human tumor tissue, magnetic cell sorting and cDNA amplification less than 500 CSCs were required for transcriptome analyses. Besides overexpressing genes involved in maintenance of stemness, the CSCs showed higher levels of genes that drive the PI3K pathway, including EGFR, HB-EGF, PDGFRA/B, PDGF, MET, PIK3CA, PIK3R1 and PIK3R2. This suggests that, in CSCs of ERα-positive breast cancer, the PI3K pathway which is involved in endocrine resistance is hyperactive.

MeSH terms

  • Breast Neoplasms / enzymology
  • Breast Neoplasms / pathology*
  • CD24 Antigen / analysis
  • Carcinoma, Ductal, Breast / enzymology
  • Carcinoma, Ductal, Breast / pathology*
  • Estrogen Receptor alpha / analysis
  • Estrogens*
  • Female
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Hyaluronan Receptors / analysis
  • Immunomagnetic Separation
  • Immunophenotyping
  • Isoenzymes / physiology
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Neoplasms, Hormone-Dependent / enzymology
  • Neoplasms, Hormone-Dependent / pathology*
  • Neoplastic Stem Cells / chemistry
  • Neoplastic Stem Cells / enzymology
  • Neoplastic Stem Cells / pathology
  • Nucleic Acid Amplification Techniques / methods*
  • Phosphatidylinositol 3-Kinases / physiology*
  • RNA, Messenger / genetics
  • RNA, Neoplasm / genetics
  • Sensitivity and Specificity
  • Transcriptome

Substances

  • CD24 Antigen
  • CD24 protein, human
  • CD44 protein, human
  • ESR1 protein, human
  • Estrogen Receptor alpha
  • Estrogens
  • Hyaluronan Receptors
  • Isoenzymes
  • Neoplasm Proteins
  • RNA, Messenger
  • RNA, Neoplasm