Effect of specific short hairpin RNA of nucleostemin on differentiation antigens and biological properties of HL-60 cells
- VernacularTitle:核干细胞因子特异性短发夹状RNA对HL-60细胞分化抗原和生物学性质的影响
- Author:
Baohong YUE
- Publication Type:Journal Article
- Keywords:
nucleostemin;
shRNA;
differentiation antigen;
biological property;
proliferation;
differentiation;
HL-60 cell
- From:Journal of Third Military Medical University
2003;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the role of the specific short hairpin RNA(NS-shRNA) of nucleostemin in inhibiting the expression of nucleostemin(NS) gene and evaluate the effect of NS-shRNA on differentiation antigen and biological properties of HL-60 cells,so as to explore the relationship between the biological functions of NS and acute leukemia,the potential perspective of NS gene therapy with RNA interference(RNAi).Methods HL60 cells were directly transfected NS-shRNA by using special transfection reagent.After 48 h,the inhibition rate of NS-shRNA to NS gene was detected by RTPCR,the proliferation ability of HL-60 cells was detected by MTT,the differentiation antigens were assayed by flow cytometry(FCM),the morphologic peculiarities such as cell shape,growth condition were observed,and the volume and granularity of HL-60 cells were analyzed by blood cell analyzer.Results The expression of NS gene were significantly inhibited by both two NS-shRNA,the inhibition rate were 37.82% and 71.88%,respectively.The significant inhibition effect of NS-shRNA to the proliferation of HL-60 cells existed in a time-dependent and concentration-dependent manner,and the best time was 48 h,the best concentration was 10 nmol/L.The change of differentiation antigen included increase of CD11b,CD33,CD14,CD64,HLA-DR and decrease of CD38,indicating continuous maturation of HL-60 cells to granulocytes and redifferentiation trend to monocytes.The aggregation of cells declined and the cell fragments increased.Myeloperoxidase(MPO) and ?-naphthol acetate esterase(?-NAE) activity increased after NS-shRNA-2 transfection.Furthermore,some HL-60 cells changed from round to fusiform shape even to pseudopod.The cells of small volume and karyorrhexis increased. Conclusion Through blocking the expression of NS gene,NS-shRNA can inhibit the cell proliferation and induce the differentiation of HL-60 cells,weaken malignant degree of HL-60 cells and trigger cell apoptosis.NS-shRNA is of clinical potential in gene therapy for acute leukemia.
