An immunofluorescent laser confocal scanning microscopical study. Mod Pathol 10:91-97īall LM, Pope J, Howard CV et al (1994) PCNA Ki-67 dissociation in childhood acute lymphoblastic leukaemia. O’Reilly PE, Raab SS, Niemann TH, et al (1197) p53, proliferating cell nuclear antigen, and Ki-67 expression in extrauterine leiomyosarcomas. J Cell Sci 116:3051–3060ĭel Giglio A, O’Brien S, Ford RJ Jr et al (1993) Proliferating cell nuclear antigen (PCNA) expression in chronic lymphocytic leukemia (CLL). Maga G, Hubscher U (2003) Proliferating cell nuclear antigen (PCNA): a dancer with many partner. BIOforum Int 5:259–261īruchova H, Borovanova T, Klamova H, Brdicka R (2002) Gene expression profiling in chronic myeloid leukemia patients treated with hydroxyurea. Cas Lek Cesk 139:655–659īruchova H, Borovanova T, Brdicka R (2001) Gene expression in patients with chronic myeloid leukemia. According to our results, nucleofection appears to be the only suitable non-viral method for siRNA delivery into the hard-to-transfect CML primary cells.īruchova H, Klamova H, Brdicka R (2000) Gene expression in chronic myeloid leukemia patients at time of diagnosis. Gene expressions ranged 22–37% that remained from original levels. In the study we obtained the best transfection efficiency using nucleofector technology. Electroporation achieved better results (suppression to 63%) but it was associated with high degree of cell death (more than 60%). Chemical transfection reagents (Oligofectamine, Metafectene, siPORT Amine) commonly used to transfect siRNAs in CML cell lines showed very low siRNA delivery in CML primary cells-mRNA levels decreased at the most to 76%. Chemically synthesized siRNAs against mentioned genes were transfected into the cells and level of knockdown was determined by real time RT-PCR. Using fluorescein-labeled siRNAs we systematically tested a variety of physical and chemical non-vector based transfection methods in order to evaluate which of them gave the most suitable transfer.
One of the crucial requirements for this purpose is a high efficiency of siRNA delivery into CML primary cells. We suppose that the genes may be implicated in the disease development and a siRNA-suppression can elucidate their functions in leukemogenesis. Our array-based analyses of gene expression in patients with chronic myeloid leukemia (CML) revealed several genes (MMP8, MMP9, PCNA, JNK2, MAPK p38) with significant increased expression. Development of array methods contributes to elucidation of many genes expressed during oncogenesis.
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