Abstract
Cell-based arrays offer powerful tools for genomics/proteomics and drug discovery, and are widely applicable for most cell lines. However, it is challenging to apply cell-based arrays for in vitro diagnosis due to limited amount of patient samples. Here, we utilized and demonstrated microfluidic image cytometry (MIC), capable of quantitative, single-cell profiling of multiple signaling molecules using only 300-3,000 cells from clinical brain tumor specimens for in vitro molecular diagnosis. First, we characterized the PI3K/AKT/mTOR pathway, which is often over-activated in the brain tumors, in U87 brain tumor cell lines by measuring EGFR, PTEN, pAKT, and pS6 with a MIC platform, and applied this measurement to clinical brain tumor specimens. In conjunction with statistical analysis, we were able to characterize extensive inter- and intra-tumoral molecular heterogeneity.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Brain Neoplasms / metabolism
-
Brain Neoplasms / pathology
-
Cell Line, Tumor
-
ErbB Receptors / analysis
-
ErbB Receptors / metabolism
-
Glioblastoma / metabolism
-
Glioblastoma / pathology
-
Humans
-
Image Cytometry / methods*
-
Immunohistochemistry
-
Intracellular Signaling Peptides and Proteins / metabolism
-
Microfluidic Analytical Techniques / instrumentation
-
Microfluidic Analytical Techniques / methods*
-
Microscopy, Fluorescence
-
PTEN Phosphohydrolase / analysis
-
PTEN Phosphohydrolase / metabolism
-
Phosphatidylinositol 3-Kinases / metabolism
-
Protein Serine-Threonine Kinases / metabolism
-
Proto-Oncogene Proteins c-akt / analysis
-
Proto-Oncogene Proteins c-akt / metabolism
-
Reproducibility of Results
-
Ribosomal Protein S6 Kinases / analysis
-
Ribosomal Protein S6 Kinases / metabolism
-
Tissue Array Analysis / methods
Substances
-
Intracellular Signaling Peptides and Proteins
-
EGFR protein, human
-
ErbB Receptors
-
Protein Serine-Threonine Kinases
-
Proto-Oncogene Proteins c-akt
-
Ribosomal Protein S6 Kinases
-
PTEN Phosphohydrolase
-
PTEN protein, human