Unit:
Τομέας Βασικών ΕπιστημώνLibrary of the School of Health Sciences
Author:
Δημακάκος Ανδρέας
Dissertation committee:
Αναπληρώτρια καθηγήτρια Παναγούλα Αγγελογιάννη
Original Title:
Μελέτη της αποσιώπησης των εξωνίων 5 και 6 του ανθρώπινου γονιδίου IGF-1 σε ανθρώπινες κυτταρικές σειρές
Summary:
Purpose: To determine if the IGF-1 Ea, Eb, Ec peptides of the IGF-1 gene
possess biological significance for the cell, aside from that of the mature
IGF-1 peptide, which is currently considered the basic biologically active
molecule. Having that in mind, a methodology was assembled to deactivate exons
5 and 6 of the IGF-1 gene, which will lead to the cessation of Ea, Eb, Ec
peptide production.
Methods: A plasmid construct was produced, to replace exon 5 with an antibiotic
sequence and introduce stop codons in all reading frames before exon 6, through
homologous recombination. The plasmid construct was transfected in PC-3
prostate cancer cells, and afterwards, the newly modified cell line was
characterized, and emphasis was given to characteristics that differed from the
wild type PC-3 cells.
Results: The PC-3 cells that successfully received the plasmid construct (EDOKO
PC-E CELLS) displayed lowered mobility and proliferation rate, while their size
was increased. They also displayed G2/M cell cycle arrest, total loss of the
cdc25b factor and senescence. However, when EDOKO PC-3 cells were inserted
subcutaneously in SCID mice for 5 weeks, they displayed greater tumorigenic
capabilities than the MOCK PC-3 cells, which were transfected with a
non-modified plasmid construct, as control samples.
Keywords:
Silencing, Insulin-like growth factors, Prostate cancer, Cell-cycle arrest, Senescence
Number of references:
164
File:
File access is restricted.
document.pdf
4 MB
File access is restricted.
