Review Article

Targeting the Epidermal Growth Factor Receptor in Epithelial Ovarian Cancer: Current Knowledge and Future Challenges

Table 1

Summary of assays used in detecting EGFR in vitro and in vivo. Aside from high-throughput methods (such as cDNA arrays, comparative genomic hybridization, and reverse phase protein arrays) and xenograft tumor assays, more broadly encompassing biological methods such as assays for invasion, migration, or gene knockouts have been excluded. cDNA: complementary DNA; PCR: polymerase chain reaction.

EGFR assay methodAssay outputPerformed in ovarian cancer?Platform for ovarian cancerReferences for ovarian cancer

cDNA ArrayDetection of mRNA levels of various genesYe Patient tissue, Human cell lines[172]
Comparative Genomic HybridizationDetection of copy number changes in chromosomesYe Patient tissue, Human cell lines [173]
Chromatin ImmunoprecipitationDetection of stable protein-DNA associationsNo
Coimmunoprecipitation + Western blottingDetection of stable protein-protein associationsNo
CrystallographyDetermination of entire structure or portions of molecule; interacting moleculesNo
Enzyme-linked Immunosorbent AssayDetermination of amount of protein in sampleYesPatient tissue[174]
Fluorescence/ Chromogenic in situ HybridizationDetermination of gene copy numberYesPatient tissue [3, 6, 35, 36]
Flow Cytometry/ Fluorescence-Activated Cell SortingDetermination of protein levels at cell surfaceYesPatient tissue, Human cell lines [175179]
Immuno-histochemistry/ Immunocyto-chemistry/ Immunofluorescence (includes Tissue Microarrays)Determination of presence, location, or amount of protein in tissue/cellYesPatient tissue, Patient effusions, Human cell lines [4, 5, 3537, 40, 41, 43, 46, 97, 117, 123, 178, 180195]
In vitro Kinase AssayMeasurement of intrinsic kinase activityNo
Mass Spectrometry after Protein Enrichment /Purification (e.g., Immunoprecipitation, Chromatographic Separation, Baculovirus Expression)Detection of protein modification sites (e.g., phosphorylation, glycosylation); changes in protein levels or proteomic profiles, protein-protein complexesNo
Microscopic Techniques (e.g., Confocal)Determination of presence, location, or amount of protein in cellNo
Mulitplex Antibody Arrays (Solid Phase or Bead Based)Detection of multiple molecules (usually proteins) of interestYe Patient serum, Human cell lines [196, 197]
Northern BlottingDetermination of steady-state RNA levelsYesPatient tissue, Human cell lines [43, 186, 193, 198, 199]
PCR + DNA analysis (e.g., Sequencing, Restriction Fragment Length Polymorphisms, Denaturing Gradient Gel Electrophoresis)Detection of known mutations/ polymorphismsYesPatient tissue, Human cell lines [6, 3537, 117, 130, 187, 200]
Quantitative PCRMeasurement of RNA levels of interestYesHuman cell lines [39, 174, 201]
Radioligand Binding/ RadioimmunoassayEstimation of number of receptors; determination of ligand or agonist/ antagonist binding kineticsYesPatient tissue, Patient effusions, Human cell lines [4245, 199]
Reverse Phase Protein ArrayDetermination of levels of several proteins and protein modifications of interestYesPatient tissue, Patient effusions [202, 203]
Reverse Transcription-PCR + Southern BlottingDetermination of mRNA levelsYesHuman cell lines, Rat cell lines [198, 204]
Southern BlottingDetection of gene of interestYesRat cell lines [198]
Tryptic Digests + Peptide Resolution (e.g., Reverse Phase High Performance Liquid Chromatography)Determination of phosphorylation sites of proteinNo
Western BlottingDetermination of protein abundance, protein-associated modifications (e.g., phosphorylation, cleavage, ubiquitination)YesPatient tissue, Human cell lines [38, 39, 46, 4850, 56, 147, 175, 177, 178, 181, 186, 196, 200, 201, 204212]
Xenograft TumorsDetermination of effect of gene/cell perturbation on tumor growthYesHuman and mouse cell lines [47, 49, 147, 178, 213219]

GFR was detected and reported, but samples were not necessarily preselected for alteration of EGFR sequence, expression, or activity.