%0 Journal Article
%T Efficient RNA interference in patients' acute lymphoblastic leukemia cells amplified as xenografts in mice
%A Ines H£¿fig
%A Harald Ehrhardt
%A Irmela Jeremias
%J Cell Communication and Signaling
%D 2012
%I BioMed Central
%R 10.1186/1478-811x-10-8
%X We established delivering siRNA into these cells without affecting cell viability. Knockdown of single or multiple genes reduced constitutive or induced protein expression accompanied by marked signaling alterations.Our novel technique allows using patient-derived tumor cells instead of cell lines for signaling studies in leukemia.Characterization of intracellular signaling mechanisms is crucial for the understanding of tumor development and for the design of novel strategies in anti-tumor therapy. For practical reasons, signaling studies are mainly performed in cell lines established from human tumors decades ago and are prone to non-representative mutations [1,2]. Example given, more than 50% of ALL cell lines inherit mutations of p53, while less than 5% of primary pediatric samples at initial diagnosis do [3-5].To overcome these limitations, several groups had successfully transfected primary leukemia cells. Best results were obtained in samples from adult patients with chronic leukemias [6-8]. In contrast, in acute leukemias, rapid onset of therapy impedes repetitive sampling and reliable results of molecular experiments in primary cells. In pediatric leukemias, small sample volumes generally disable molecular work on these primary cells. For the transfection of childhood ALL samples by nucleofection, so far transgene expression was studied in detail rendering varying results between 1% and 62.3% [9].Acute leukemia samples can be amplified in severely immuno-compromised mice. The xenograft mouse model of acute leukemia has been characterized to enable frequent engraftment with little genetic and phenotypic alterations upon passaging through mice [10]. The regular detection of CD surface markers revealed stable phenotypes over several passages [11].Here, we describe a new method which will routinely allow repetitive and reliable molecular signaling studies on patient-derived childhood ALL cells amplified in NOD/SCID mice. The aim was to establish a suitable trans
%U http://www.biosignaling.com/content/10/1/8