Determination of PML-RARalpha gene expression by using the RT-qPCR technique from blood cells RNA in murine model of acute promyelocytic leucemia

Our goal was to develop an accurate, quick and affordable method for following up the progression of acute promyelocytic leucemia in murine model. We took advantage of a developed system of reverse transkriptase quantitative polymerase chain reaction (RT-qPCR) performinng human bone marrow samples to develop a quantitative method of measurement of PML-RARalpha transcripts for murine model. PML-RARalpha transcripts are hallmarks of acute promyelocytic leucemia (APL) with t(15;17) translocation. For measuring bcr1 transcripts, RNA was isolated from murine blood. We present here a method for quantification of t(15;17) transcript bcr1, which is developed in myeloid cells of transgenic mice and then is injected into FVB/N mice. The expression of PML-RARalpha transcripts is normalized using the housekeeping 18S ribosomal subunit gene. We have quantitative data for three healthy mice (FVB/N) and for six ill mice (APL), which were injected 104, 103 or 102 blasts. We show negative expression in FVB/N mice and different copy numbers for APL mice. There is not shown a complete correlation between the amount of injected cells and the amount of transkript copies