Inhibitory effect of RNA interference targeting GFI-1 on the proliferation of atypical chronic myelogenous leukemia NT1 cells
10.3760/cma.j.issn.0253-3766.2016.08.003
- VernacularTitle:靶向沉默独立生长因子1基因对不典型慢性髓细胞白血病细胞生长和增殖的影响
- Author:
Xi YANG
1
;
Hong LIU
;
Zenghua LIN
;
Juan QIAN
;
Xinrun XU
Author Information
1. 226000,南通大学附属医院血液科
- Keywords:
Leukemia,myeloid;
Cell;
GFI-1;
Cell proliferation;
RNA,small interfering
- From:
Chinese Journal of Oncology
2016;38(8):572-577
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effects of RNA interference targeting GFI?1 on growth and proliferation of atypical chronic myelogenous leukemia ( aCML) NT1 cells. Methods NT1 cells were transfected with PBS and liposome complex ( vehicle group) , scrambled siRNA and liposome complex ( negative control, NC group ) , and GFI?1 siRNA and liposome complex ( GFI?1 siRNA group ) , respectively. Real?time quantitative RT?PCR ( qRT?PCR) and Western blot were performed to examine the expression levels of GFI?1 mRNA and protein, respectively. The proliferation abilities of NT1 cells of the three groups were evaluated by MTT assay. The cell cycle in cells of the three groups was analyzed by flow cytometry. Moreover, nude mouse xenograft model was used to detect the tumor formation ability in the three group cells. Results Quantitative real?time PCR data showed that the expression level of GFI?1 mRNA in GFI?1 siRNA group was significantly lower than those of NC group and vehicle group [(0.367±0.017) vs. (0.918±0.006) and (1.010±0.005), respectively, (P<0.05)]. Western blot results showed that the GFI?1 protein expression level in the GFI?1 siRNA group was also significantly reduced, compared with those of the NC group and vehicle group ( P<0.05 for both) . From MTT assay data, the absorbance value of NT1 cells in the GFI?1 siRNA group (0.667±0.059) was significantly lower than those of the NC group (1.096±0.049) and vehicle group (1.193±0.064, P=0.023). Flow cytometry data showed that sub?G1 and G0/G1 phase proportions of the GFI?1 siRNA group were significantly higher than those of the NC and vehicle groups [ sub?G1: (8.2±2.5)% vs. (1.9±1.3)% and (2.0±3.6)%, respectively, (P<0.05);G0/G1:(66.7±3.8)% vs. (53.3±4.5)% and (48.6±3.2)%, respectively, (P<0.05)]. Furthermore, the tumor weight in the GFI?1 siRNA group [(0.37±0.02) g] was significantly lower than those in the NC group [(0.83±0.06) g] and vehicle group [(0.92±0.04) g] (P<0.05). Conclusions RNA interference targeting GFI?1 inhibits the growth and proliferation of NT1 cells, which may provide a new therapeutic target for atypical chronic myelogenous leukemia.
