Home > Browse Issues > Vol.34 No.9
Effect of Nucleophosmin (NPM1) Mutations on the Differentiation of K562 Leukemic Cells
Tan Shi, Zhang Huijuan, Wang Juan, Chen Shana, Qin Fengxian, Chen Xianchun, Zhang Ling*
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China
Abstract: Nucleophosmin (NPM1) mutations play important roles in the leukemogenesis， whereas the relationship between NPM1 mutations and the differentiation block of leukemic cells have not yet been elucidated. To explore the effect of NPM1 mutations on the in vitro differentiation of leukemic cells， the plasmid pEGFPC1- NPM1-mA carrying NPM1 mutation A (NPM1-mA) was transfected into K562 cells， and the leukemic cells with stably expressed NPM1-mA protein (K562 mA) were established. The K562 cells transfected with pEGFPC1- NPM1-wt (K562 wt)， pEGFP-C1 (K562 C1) and the untreated cells (K562) were used as control. After induction with phorbol-12-myristate-13-acetate (PMA)， the morphological changes of cells were observed under light microscope following Wright-Giemsa staining and the differentiation rate was estimated. Adherent cells were examined by phase contrast light microscopy. The expressions of surface marker CD41 were analyzed by flow cytometry. Compared with the control groups， the differentiation rates and the percentage of adherent cells in K562 mA group decreased obviously after PMA treatment for 72 h (P<0.05). Meanwhile， the expression of CD41 was inhibited significantly in K562 mA group (P<0.01). Our data indicate that NPM1 mutations block differentiation of K562 leukemic cells in vitro.
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