Development of cell arrays - cell line screening on immunocytochemistry base assessed by new software COLGRAPH

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