Konference: 2007 3. ročník Dny diagnostické, prediktivní a experimentální onkologie
Kategorie: Onkologická diagnostika
Téma: Postery
Číslo abstraktu: 008p
Autoři: Mgr. Pavla Hublarová; Ing. Dana Knoflíčková; Zina Hanzelková; Y. Baturko; MUDr. Rudolf Nenutil, CSc.; RNDr. Bořivoj Vojtěšek, DrSc.
Cell arrays are new approach to the standardization of
immunohistochemical methods (IHC). They are suitable for i)
evaluation of IHC, ii) characterization of antibody, iii) proteomic
research and iv) are ideal tool for extensive studies carried on
cell lines. Great advantages of this method are low material usage,
huge volume of highly reproducible data assessed by computer
analysis with defined colour intensity levels and possibility of
comparing 23 different specimens of cell lines in one experiment.
Primarily, cell suspensions are fixed in formalin, embedded in
agarose and than in paraffin to form blocks. Subsequently,
cylinders (1 mm in diameter) are punctuated from the each block,
and placed in distinct positions in cell array. Complete array is
than sliced by microtom and thawed to SuperFrost+ glass. The
purpose of our cell array development was to overcome the problems
of classical immunocy-tochemistry on formalin-fixed
paraffin-embedded material and to improve the testing of newly
developed antibodies. By this methodical approach, we analysed the
expression of p53 and MDM2 proteins with antibody DO-1 and 2A9,
respectively, in breast cancer cell lines using newly designed
software for automatic assessing IHC - ColorGraphicAnalysis
(COLGRAPH). We quantified p53 expression and determined p53 status
in cancer cell lines on the basis of comparing histoscores of
proteins p53 and MDM2. We successfully pointed up a distribution of
wild type and mutant p53 cell lines according to homogeneity of p53
expression and positivity of p53 IHC reaction. We developed a new
methodological approach to IHC that is full of flexibility. The
cell array can be designed according to acquired attributes like
cell origin, protein expression, studied treatment etc. In the
combination with new software ColorGraphicAnalysis (COLGRAPH),
above meant method gives us a powerful tool for sample screening in
extensive studies. On the basis of our promising results, this
method is going to have rich possibilities of application in
research and diagnostics fields in future.
This work was supported by grants IGA MZ CR NR /8338-3/ 2005, MSMT
LC06035 and MZ0 00209805.
Datum přednesení příspěvku: 28. 11. 2007
