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ORIGINAL ARTICLES  TRENDS IN MOLECULAR DIAGNOSIS AND THERAPY OF β-THALASSEMIA AND SICKLE CELL ANEMIA


Minerva Biotecnologica 2003 Giugno;15(2):153-60

lingua: Inglese

Use of mac­ro­ar­ray tech­nol­o­gy to ­study the ­effects of DNA-bind­ing ­drugs on ­gene expres­sion pro­file of eryth­roid-­induced ­human leu­kem­ic K562 ­cells

Mischiati C. 1, Sereni A. 1, Gambari R. 1, 2

1 Department of Biochemistry and Molecular Biology, University of Ferrara, Ferrara, Italy;
2 Biotechnology Center, University of Ferrara, Ferrara, Italy


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In ­this ­paper, we ­review ­recent ­data ­obtained in our labor­a­to­ry, ­aimed to deter­mine the ­effects of DNA-bind­ing mole­cules on ­gene expres­sion pro­file and their abil­ity to ­induce dif­fe­ren­ti­a­tion in K562 ­cells and HbF pro­duc­tion in eryth­roid pre­cur­sor ­cells iso­lat­ed ­from the periph­er­al ­blood of nor­mal ­donors as ­well as tha­las­se­mia ­patients. Here, we ­focused the atten­tion on the molec­u­lar ­effects of tal­li­mus­tine, a DNA bind­ing ­drug ­with selec­tiv­ity for AT sequenc­es ­that was dem­on­strat­ed to ­induce eryth­roid dif­fe­ren­ti­a­tion of ­human K562 leu­ke­mia ­cells as ­well as HbF pro­duc­tion in eryth­roid pre­cur­sor ­cells. By ­using mac­ro­ar­ray tech­nol­o­gy, we iden­ti­fied up- and ­down-reg­u­lat­ed ­genes fol­low­ing treat­ment of K562 ­cells ­with tal­li­mus­tine. These ­results ­were con­firmed by ­reverse-tran­scrip­tion poly­me­rase-­chain reac­tion (RT-PCR) anal­y­sis. Molecular anal­y­sis of the pro­mot­er of ­these ­genes, of the sequenc­es of the mRNA(s), of the struc­ture of the encod­ed pro­tein ­will ­allow to ­design ­decoy ODN, anti­sense DNA or RNA, pep­tides or mono­clo­nal anti­bod­ies expect­ed to mim­ick the bio­log­i­cal ­effects of the ­employed induc­er, lim­it­ing at the ­same ­time pos­sible ­side ­effects.

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