Effect and possible mechanism of HnRNP E2 decoy RNA on proliferation of K562 leukemia cells

2006 
BACKGROUND & OBJECTIVE: The abnormal expression of poly(rC)-binding protein E2 (hnRNP E2) induced by BCR/ABL plays an important role in blast crisis of chronic myeloid leukemia (CML). The present study was to investigate the effect of hnRNP E2 decoy RNA on the cell proliferation in K562 leukemia cells, and further elucidate the possible underlying mechanisms. METHODS: Decoy hnRNP E2 plasmid was constructed and transfected into K562 cells using cationic liposome. Stably transfected cells were selected with G418. The cell proliferation rate was determined by cell growth curve using trypan blue staining, and the cell cycle was analyzed by flow cytometry. The changes of CCAAT/enhancer-binding protein alpha (C/EBP alpha) and c-Myc gene expression were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. RESULTS: The proliferation rate of stably transfected K562 cells was inhibited by (62.73+/-12.92)%. The cell cycle was arrested at S phase [stably transfected K562 cell group: (55.59+/-4.67)%, control group: (44.70+/-4.21)%, P<0.05]; C/EBPalpha mRNA level remained unchanged. However the 42 ku-C/EBPalpha protein expression was elevated by (49.72+/-5.58)%; c-Myc mRNA and protein expression was inhibited by (58.27+/-7.23)% and (57.26+/-6.52)%, respectively. CONCLUSION: HnRNP E2 decoy RNA could inhibit the proliferation of K562 cells, and this may be caused by the blockage of the binding between hnRNP E2 and C/EBPalpha mRNA and subsequent elevation of 42 ku-C/EBPalpha by decoy RNA.
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