Zbornik - Društvo genetičara Srbije

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196 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE V-Usm-13 Subotica, 30. novembar - 4. decembar 2004. ANALIZA EKSPRESIJE GENA PRIMENOM NERADIOAKTIVNE RNK-RNK IN SITU HIBRIDIZACIJE Gordana Nikèeviæ, Danijela Drakuliæ i Milena Stevanoviæ Institut za molekularnu genetiku and genetièko inenjerstvo, Beograd RNK-RNK in situ hibridizacija je pogodna metoda za izuèavanje specifiène ekspresije gena u tkivima i æelijama, kojom je omoguæena vizualizacija obeleene antisens RNK probe hibridizovane za specifiènu iRNK. U ovom saopštenju biæe prikazano detektovanje ekspresije SOX gena u tumorskim æelijskim linijama primenom neradioaktivne RNK-RNK in situ hibridizacije koristeæi RNK probe obeleene biotinom ili digoksigeninom. Koristeæi ovaj pristup potvrdili smo rezultate dobijene Northern blot analizom, tj. pokazano je prisustvo SOX2 iRNK u NT2/D1 i SOX14 iRNK u HepG2 æelijama. Cilj ovog rada bio je uspostavljanje RNK-RNK in situ hibridizacije na in vitro kultivisanim æelijama da bi metodu primenili pri prouèavanju ekspresije SOX gena u razlièitim normalnim i tumorskim tkivima. GENE EXPRESSION ANALYSIS BY NON-RADIOACTIVE RNA-RNA IN SITU HYBRIDIZATION TECHNIQUES RNA-RNA in situ hybridization is reliable method for studying tissue and cell specific gene expression, which enables visualization of labeled antisense RNA probe hybridized to specific mRNA. In this study we have employed non-radioactive RNA-RNA in situ hybridization using biotin- or digoxigenin-labeled RNA probes in order to detect SOX gene expression in carcinoma cell lines. By applying this approach we confirmed results obtained by Northern blot analysis, where presence of SOX2 mRNA in NT2/D1 and SOX14 mRNA in HepG2 cells has been shown. Our aim was to set up RNA-RNA in situ hybridization method in in vitro cultured cells in order to further analyze SOX gene expression on various normal and cancer tissues.
V-Usm-14 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 197 Subotica, 30. novembar - 4. decembar 2004. DETEKCIJA MINIMALNE REZIDUALNE BOLESTI KOD BOLESNIKA SA HRONIÈNOM MIJELOIDNOM LEUKEMIJOM (CML) LEÈENIH IMATINIB MESYLATOM Anka Radoviæ, Biljana Todoriæ-ivanoviæ, Milica Strnad, Dragana Stamatoviæ, Koviljka Krtolica 1 i Z. Magiæ 1 Vojnomedicinska Akademija, Beograd 1 Institut za nuklearne nauke «Vinèa», Beograd Detekcija i praæenje minimalne rezidualne bolesti (MRB), zaostalih malignih æelija nakon terapije, postalo je jedno od najvanijih zadataka kad su u pitanju bolesnici sa CML leèeni imatinib mesylatom. Postizanje kompletne hematološke i citogenetske remisije kod odreðenog broja bolesnika sa CML a zatim relaps bolesti objašnjavaju se upravo postojanjem malog broja malignih æelija rezistentnih na primenjenu terapiju. Zbog toga njihovo otkrivanje daje vanu informaciju o efektu terapije kao i moguænost da terapeut pravovremeno odreaguje pre nego što doðe do relapsa bolesti. RT-PCR tehnika se pokazala kao najsenzitivnija, omoguæujuæi detekciju jedne maligne æelije na 10 5-106normalnih. Kao pozitivni marker za detekciju MRB koristi se BCR-ABL fuzioni gen koji ima kljuènu ulogu u patogenezi CML. Detekcija prisustva minimalne rezidualne bolesti bolesnika sa CML koji su postigli kompletan citogenetski odgovor (0% æelija sa Philadelfija hromozomom) nakon primenjene terapije imatinib mesylatom. Prisustvo rezidualnih æelija koje eksprimiraju BCR-ABL himerni gen analizirano je u uzorcima periferne krvi kod pet bolesnika (od dvadeset na imatinib terapiji) koji su postigli kompletan citogenetski odgovor. Detekcija je vršena RT-PCR i «nested» RT-PCR tehnikom. Kod tri bolesnika RT-PCR tehnikom detektovano je prisustvo rearanmana b3a2 tipa. Kod dva bolesnika je po dobijanju negativnog rezultata nakon RT-PCR tehnike primenjena «nested» RT-PCRmetoda. Ovom metodom utvrðeno je prisustvo BCR-ABL rearanmana i to kod jednog bolesnika b3a2 tip a kod drugog b2a2 tip. DETECTION OF MINIMAL RESIDUAL DISEASE IN PATIENTS WITH CHRONIC MYELOGENOUS LEUKEMIA (CML) TREATED WITH IMATINIB MESYLATE Detection and monitoring of minimal residual disease (MRD), malignant cells remained after the therapy, has become one of the most important tasks when it comes to CML patients treated with imatinib mesylate. The fact that certain number of patients suffering from CML first achieve complete hematologic and cytogenetic remission which is afterwards followed by relapse of disease could actually be clarified through the existence of small number of malignant cells that are resistant to the applied therapy. Therefore, detection of these cells will provide important information on the effect of therapy as well as offer a therapist an opportunity to react promptly before the relapse of disease takes place. RT-PCR technique proved to be the most sensitive one enabling the detection of one malignant cell in 10 5 -10 6 normal ones. BCR-ABL fusion gene that has a crucial role in pathogenesis of CML is used as positive marker for detection of MRD. Detection of the presence of minimal residual disease in CML patients who after the application of imatinib mesylate therapy reached complete cytogenetic response (0%cells with Philadelphia chromosome). Presence of residual cells that express BCR-ABL chimerical gene was analysed in the samples of peripheral blood from five patients (out of twenty undergoing imatinib therapy) who had reached complete cytogenetic response. Detection was performed by the means of RT-PCR and nested RT-PCR technique. In three patients b3a2 rearrangement type was detected by RT-PCR tehnique. In two remaining patients after negative RT-PCR results nested RT-PCR was applied. By this method BCR-ABL rearrangement was detected in both patients, one of them having b3a2 type and the other b2a2 type.
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DRUŠTVO GENETIČARA SRBIJE SERBIAN
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Zbornik - Društvo genetičara Srbije

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