Unit:
Κατεύθυνση Καρκίνος Πνεύμονα: Σύγχρονη Κλινικοεργαστηριακή Προσέγγιση και ΈρευναLibrary of the School of Health Sciences
Supervisors info:
Καθηγητής Κωνσταντίνος Συρίγος
Original Title:
In vitro ανίχνευση και ποσοτική μέτρηση του συμπλέγματος του επιδερμοειδούς αυξητικού παράγοντα (Epidermal Growth Factor Receptor - EGFR)
Summary:
Epidermal Growth Factor Receptor (EGFR) is a therapeutic target in Non-Small
Cell Lung Cancer (NSCLC) for first-line treatment in EGFR mutant adenocarcinoma
patients. The presence of activating mutations is associated with better
overall survival compared to patients with wild type receptor, irrespective of
stage and treatment. However, the data about the prognostic role of overall
EGFR expression measured by immunohistochemistry (IHC) in NSCLC are conflicting
with most studies showing no prognostic effect. EGFR gene copy number
assessment by fluorescent in situ hybridization (FISH) has also failed to show
a consistent association between increased EGFR gene copy number and prognosis.
Proximity ligation assay (PLA) is a method to detect functional signaling
associated protein complexes. In previous studies, PLA has been used to detect
the interaction between EGFR and growth factor receptor-bound protein 2 (GRB2).
GRB2 is an adaptor protein which binds to the phosphorylated residues of active
receptor. The interaction between EGFR and GRB2 correlates with active EGFR
signaling and leads to activation of MAPK/ERK pathway.
A PLA developed to detect EGFR:GRB2 interaction was measured by quantitative
immunofluorescence (QIF) using AQUA(Automated Quantitative Immunofluorescence)
technology. EGFR pathway activation was assessed in 6 cell lines with different
mutation status and overall EGFR expression.PLA measured by AQUA was found to
be correlated to p-EGFR as measured by Western Blot in the 6 cell lines.
Additionally, PLA measured by AQUA subjectively and reproducibly assessed
EGFR:GRB2 interaction in serial cuts and different patient tumor blocks.
Keywords:
Complex, Receptor, Epidermal, Protein, Immunofluorescence
