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背景:细胞的多倍性是重要的生物学现象,不仅影响组织器官和造血系统的正常生理功能,而且是某些应激条件下代偿性的表现,和发生某些病理过程的基础,尤其与恶性肿瘤的发生有着密切的关系。目的:优化SP600125诱导Dami细胞多倍体化模型,探讨姜黄素对此多倍体化逆转作用的机制。方法:将不同浓度SP600125(15,30,60mmol/L)分别加入细胞悬液中,诱导Dami细胞多倍体化,确定最佳诱导时间和浓度;将5,10,20mmol/L姜黄素分别加入含有SP600125的细胞悬液中,培养72h收集细胞。AnnexinV-PI染色法的流式细胞术检测细胞倍性的变化、Westernblot法检测细胞周期相关蛋白的变化。结果与结论:30mmol/LSP600125作用Dami细胞72h,诱导的多倍体化细胞数量最多,并且cyclinD3表达最高;姜黄素明显逆转Dami细胞多倍体化,cyclinD3表达逐渐降低,存在明显的剂量依赖关系。结果显示,实验成功建立了Dami细胞的多倍体化模型,并确立了诱导多倍体化Dami细胞的SP600125最佳诱导时间和浓度72h,30mmol/L。姜黄素可能通过抑制cyclinD3的表达,逆转SP600125诱导Dami细胞多倍体化。
BACKGROUND: Polyploidy of cells is an important biological phenomenon that affects not only the normal physiological functions of tissues and organs and the hematopoietic system but also the compensatory performance under certain stress conditions and the basis for the development of certain pathological processes, in particular And the occurrence of malignant tumors are closely related. OBJECTIVE: To optimize the model of Dami cell polyploidization induced by SP600125 and to explore the mechanism of curcumin on the reversal of polyploidization. Methods: Different concentrations of SP600125 (15, 30, 60mmol / L) were added to the cell suspension to induce Dami cell polyploidy, and the optimal induction time and concentration were determined. 5,10,20mmol / L curcumin The cell suspension containing SP600125 was cultured for 72 hours to collect the cells. AnnexinV-PI staining was used to detect the change of cell ploidy. The changes of cell cycle related proteins were detected by Western blot. RESULTS AND CONCLUSION: Dami cells treated with 30mmol / L LSP600125 for 72h induced the highest number of polyploidy cells and the highest expression of cyclinD3. Curcumin reversed Dami cell polyploidization and cyclinD3 expression gradually decreased in a dose-dependent manner. The results showed that the polyploidization model of Dami cells was successfully established and the optimum induction time and concentration of SP600125 induced by polyploidized Dami cells were 72 h and 30 mmol / L. Curcumin may reverse D600 cell polyploidization by reversing the expression of cyclinD3.