Rna Interference Inhibits Bcr/Abl Fusion Protein Expression And Effect On Chronic Myelogenous Leukemia Cell K562

The effect of small interfering RNA on the Chronic myelogenous leukemia cell line K562and the expression level of p210bcr/abl were detected. The lentiviral expression vectors for the follow-up study were constructed, whicth provided a basis of the research.The cells were transfected with liposome, and the cell activation was tested with MTT assay. The cells'ability of colony formation was analyzed quantitatively using soft agar colony formation assay.The cell apoptosis was tested with Annexin-V-FITC, and the expression level of p210bcr/abl protein was measured by western blot. Whether the lentiviral expression vectors packaging were successful was detected by fluorescence microscope.The most suitable concentration of siRNA transfection was100nM.With Qiagen HiPerFect Transfect Reagent, the transfection efficiency could be50%.48h after transfection, compared with the negative control, anti-land anti-4oligo could reduce the cell activation to56.5%and57.2%.72h after transfection, the early apoptosis rate of anti-1,anti-4and negative control were8.08%,14.18%, and4.19%, the late apoptosis and death rate were15.48%,9.88%, and2.13%.48h after transfcetion, the expression of p210of anti-1and anti-4was reduced. Lentiviral vectors were packaged in HEK-293cell, and72h after transfection of plasmids,70%of cell expressed Green fluorescent.Small interfering RNA can effectively inhibit the activity of K562cells, inhibit the proliferation, induce the apoptosis, impede the cell clone formation, and decreased the expression level of p210fusion protein, which provided theory evidence for RNAi therapy of CML. The success of construction of lentiviral expression vectors, built a platform for our research, and provided a new research method.