Effect of transforming growth factor-beta1 on proliferation and apoptosis of mononuclear cells of cord blood.
- Author:
Bin SHI
1
;
Xiang-Fu ZHAO
;
Jiang-Ying CUI
;
Shen-Wu WANG
Author Information
1. Department of Hematology Fuxing Hospital, Capital Medical University, Beijing 100038, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
drug effects;
CDC2-CDC28 Kinases;
analysis;
Cell Proliferation;
drug effects;
Cells, Cultured;
Cyclin A;
analysis;
Cyclin D1;
analysis;
Cyclin-Dependent Kinase 2;
DNA;
biosynthesis;
Fetal Blood;
cytology;
Humans;
Leukocytes, Mononuclear;
drug effects;
Transforming Growth Factor beta;
pharmacology;
Transforming Growth Factor beta1
- From:
Journal of Experimental Hematology
2004;12(5):644-648
- CountryChina
- Language:Chinese
-
Abstract:
To explore the mechanism of transforming growth factor-beta1 (TGF-beta1) effect on umbilical cord blood mononuclear cells proliferation and apoptosis, 5-bromo-2'-deoxyurine (BrdU) incorporation assay was adopted to detect effect of TGF-beta1 on synthesis of DNA in cells. Western blot method was used to examine effect of TGF-beta1 on expression of cyclin A, Cyclin D1, CDK2 and CDK4 in G1 phase of cell cycle. Giemsa staining and flow cytometry (FCM) were performed to detected effect of TGF-beta1 on cell apoptosis. The results showed that (1) after culture of cells with IMDM containing 10% FBS, 10% FBS + 1 ng/ml TGF-beta1, 10% FBS + 2 ng/ml TGF-beta1 or 10% FBS + 5 ng/ml TGF-beta1 for 12 hours the OD values of TGF-beta1 group were significantly lower than control group (P <0.01); after culture for 24 hours the OD values of 1 ng/ml TGF-beta1 group had no significant difference compared with control group (P >0.05), but the OD values of 2 ng/ml and 5 ng/ml TGF-beta1 groups were significantly lower than control group (P <0.05). (2) 2 ng/ml TGF-beta1 could significantly inhibit the production of cyclin A, cyclin D1, CDK2 and CDK4, the protein levels were significantly lower than control group. (3) when the cells were co-cultured with 2 ng/ml TGF-beta1 for 12 and 24 hours, Giemsa staining and FCM detection could display typical apoptosis, the apoptosis rates were 14.42% and 31.98%, while apoptosis rate in control were 4.71% and 5.76%. It is concluded that TGF-beta1 can inhibit production of G1 cyclins and CDKs of umbilical cord blood mononuclear cells, arrest cells in the G1 phase of cell cycle and induce cell apoptosis. Thus, TGF-beta1 may be an important negative modulator in hematopoiesis.
