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目的探讨Flavopiridol(FP)联合顺铂(DDP)对人骨肉瘤细胞株(U2-OS)的增殖及凋亡的作用。方法将不同浓度的FP(0、50、100、200、400、1000nmol·L~(-1))分别与人U2-OS细胞共同培养24、48h后,采用MTT法检测U2-OS细胞增殖,以确定FP作用48h的半数抑制浓度(IC50);采用Hoechst 33258染色法观察细胞凋亡的形态学改变;采用流式细胞术测定U2-OS细胞凋亡率。然后将培养后的U2-OS细胞分为4组:空白对照组、单用FP组、单用DDP组、联合用药组(FP+DDP组),每组设5个复孔,分别加入0.1%DMSO、FP(IC50)、DDP、FP(IC50)+DDP处理,采用MTT法检测细胞增殖,流式细胞术检测细胞凋亡率和细胞周期。结果 FP对人U2-OS细胞的增殖抑制作用呈现浓度依赖性,作用48h的IC50值为340nmol·L~(-1),Hoechst33258染色后细胞出现明显的核固缩、核浓聚等凋亡改变,随着FP浓度的增加,流式细胞术检测细胞凋亡率升高。FP+DDP联合作用于人U2-OS细胞,随着时间的增加抑制率逐渐增大(P<0.05);与FP和DDP单独用药组比较,抑制率、凋亡率及周期阻滞差异均有统计学意义(P<0.05)。结论 FP可明显抑制人U2-OS细胞增殖并诱导凋亡,联合顺铂对人U2-OS细胞的增殖抑制及凋亡促进具有协同作用。
Objective To investigate the effect of Flavopiridol (FP) and cisplatin (DDP) on the proliferation and apoptosis of human osteosarcoma cell line (U2-OS). Methods Different concentrations of FP (0, 50, 100, 200, 400 and 1000 nmol·L -1) were incubated with human U2-OS cells for 24 and 48 h respectively. MTT assay was used to detect the proliferation, To determine the half-maximal inhibitory concentration (IC50) of FP for 48h. Morphological changes of apoptosis were observed by Hoechst 33258 staining. The apoptosis rate of U2-OS cells was determined by flow cytometry. The cultured U2-OS cells were divided into 4 groups: blank control group, single FP group, single DDP group and combination therapy group (FP + DDP group) DMSO, FP (IC50), DDP, FP (IC50) + DDP. The cell proliferation was detected by MTT assay. The apoptosis rate and cell cycle were detected by flow cytometry. Results FP inhibited the proliferation of human U2-OS cells in a concentration-dependent manner. The IC50 value was 340 nmol·L -1 after 48 h of treatment. The apoptotic changes of nuclei after nuclear Hoechst 33258 staining were observed. , With the increase of FP concentration, flow cytometry detected apoptosis rate increased. FP + DDP combined with U2-OS cells, with the increase of time, the inhibitory rate gradually increased (P <0.05); Compared with FP and DDP alone group, the inhibition rate, apoptosis rate and cycle retardation were Statistical significance (P <0.05). Conclusion FP can significantly inhibit human U2-OS cell proliferation and induce apoptosis. Combined with cisplatin, it has a synergistic effect on the proliferation inhibition and apoptosis promotion of human U2-OS cells.