Abstract:

BACKGROUND: Cell polyploidization is an important biological phenomenon. It not only influences the normal physiological function of tissue and hematopoietic system, but also is the compensatory performance under stress condition. It is closely related to the occurrence of some pathological processes, in particular malignant tumorigenesis.
OBJECTIVE: To optimize the Dami cell model of SP600125-induced polyploidization and to investigate the mechanism by which curcumin reversed the polyploidization.
METHODS: 15, 60, 60 μmol/mL SP600125 was added to the cell suspension to induce the polyploidization of Dami cells. The optimal induction time and concentration were determined. Then 5, 10, 20 mmol/L curcumin was added to the cell suspension containing SP600125. Cells were collected after 72 hours of culture. Cell ploidy changes were determined by flow cytometry using Annexin V-PI staining. Cell cycle-related protein expression changes were detected by Western blot method.
RESUITS AND CONCLUSION: After treated with 30 mmol/L SP600125 for 72 hours, the number of polyploid Dami cells was increased and the expression of cyclin D3 was highest. The ployploidization of Dami cells was significantly reversed by curcumin, and the expression of cyclin D3 was gradually decreased, showing a significant time- and dose-dependent manner. A Dami cell model of polyploidization was successfully established and the optical induction time and concentration for SP600125 were determined at 72 hours and 30 mmol/L. The polyploidization of Dami cells may be reversed through inhibiting cyclinD3 expression.