A analysis of the proteome of HL60 cell line induced by a new steroidal drug as a monocytic inducer
- VernacularTitle:甲磺酸甾醇类药物诱导HL60细胞分化及其蛋白质组表达差异的研究
- Author:
Weijia WANG
;
Wei TANG
;
Xiaoqin MAO
;
Zongyin QIU
- Publication Type:Journal Article
- Keywords:
drug,steroidal;
HL60 cell;
cellular differentiation;
two-dimensional electrophoresis
- From:
Chinese Pharmacological Bulletin
2003;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim The potentiality as a differentiation inducer of the new steroidal drug(NSC67657) had been studied.Then the expression differences of protein between treated and untreated HL60 cell line could be analysed.Method Firstly,the expression patterns of C/EBP alpha gene and protein were observed between treated and untreated HL60 cell line.Then the proliferation of HL60 cell line could be investigated by MTT.At the same time cellular chemical staining could be employed to investigate which direction HL60 cell line would be induced by NSC67657.Then the flow cytometry(FCM) could be employed to detect the profile of differentiation of HL60 cell line induced in different time and at different drug concentrations,by which the most suitable drug concentration and inducing time could be found. Following that,the information of cellular cycle and ultramicrostructure could be analysed by FCM and electronmicro scope,by which whether the apoptosis had happened or not under the drug treatment could also be found. Finally,the protein of these two group HL60 cell lines could be separated by modified two-dimensional electrophoresis (2-DE).Results The expression of C/EBP alpha gene and protein could be promoted under the treatment of NSC67657.Then the proliferation of HL60 cell line was inhibited significantly.From cellular chemical staining,the monocytic differentiation could be easily found and the perfect inducing time and drug concentration were defined as 10 ?mol?L-1NSC67657 and constantly inducing HL60 cell line within 5 days.The cellular number of G0~G1 was increased and hardly any apoptosis which might be happened during drug inducing ability could be seen.The protein of HL60 cell lines were separated by modified 2-DE technology.Then there were 14 protein spots which could only be found in the differentiated gels,on the other hand,20 protein spots could only be found in the undifferentiated gels,which would have been analyzed by MALDI-TOF.Conclusions HL60 cell line could be induced to monocyte by NSC67657 which could also stimulate the C/EBP? in the early stage.2-DE could separate the protein directly which expressed differentially,from which some proteins essential in cellular differentiation might be found.
