Место издания:Blackwell Publishing Inc United Kingdom
Первая страница:466
Последняя страница:466
Аннотация:The speculations about the role of MUC1 which is known to be
overexpressed in glandular cancers on metastatic potential of
cancer cells originate from the data indicating that cell malignization
correlates with the change of apical mucin MUC1 localization
to peripheral [1]. But the role of MUC1 in cancer
metastasizing is not clear yet [2-4]. The major problem is the
absence of adequate cell models. The aim of this study was to
create cell models presenting different fragments of human mucin
MUC1 extracellular domain on cell surface.
Genetic constructions were generated on the basis of plasmid
vector pEGFP-N3. These constructions contain fusion genes coding
chimeric proteins composed of different combinations of mucin
MUC1 functional domains and identification markers – FLAGepitope
located at the N-terminus and EGFP located at the C-terminus
of the chimeric proteins. The constructions were used for
stable transformation of human cancer cells HT-29. Transformants
obtained were characterized by means of flow cytometry. Low
expression level of endogenous mucin MUC1 and high expression
level of the recombinant proteins were confirmed by real-time
PCR. Microscopic examination of transformed cells confirmed
localization of fusion proteins on its plasmatic membrane. Cells
obtained are used to be a quite applicable model of MUC1-
expressing cancers and might be used for study the role of different
functional fragments of mucin MUC1 in metastasizing.
Cells obtained were tested for the ability to penetrate and
migrate through the endothelium by using an artificial system. It
was shown that the expression of fragments of mucin MUC1
extracellular domain leads to an increase in the proportion of
cells capable to overcome the endothelial barrier. Furthermore, a
fragment has been identified, the presentation of which on the
cell surface leads to the greatest increase in the proportion of
cells migrated.