Flavopiridol联合顺铂对人U2-OS细胞增殖及凋亡的影响

Effects of Flavopiridol Combined with Cisplatin on Proliferation and Apoptosis of Human Osteoblastic U2-OS Cells

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 effects of flavopiridol (FP) combined with cisplatin (DDP) on the proliferation and apoptosis of human osteoblastic U2-OS cells. Methods U2-OS cells were treated with different concentrations of FP (0,50,100,200,400 and 1000 nmol ? L_1) for 24 and 48 hours. Cell viability was measured by MTT assay,and the IC5〇 value for FP was calculated after treatment for 48 hours. The morphological changes in apoptotic cells were observed by Hoechst 33258 staining,and the apoptosis rate was assessed by flow cytometry. Furthermore,the cultured U2-OS cells were divided into four groups:0. 1% DMSO treatment group (control group),FP (IC5〇) treatment group,DDP treatment group,and FP (IC5〇)+DDP group,with five wells in each group. Cell proliferation was detected by MTT assay. Cell apoptosis and cell cycle were determined by flow cytometry. Results The inhibitory effect of FP on the proliferation of U2-0S cells was dose-dependent (P<C0. 05) ,and the IC5〇 value for FP was 340 nmol ? L_1 at 48 hours after treatment. Hoechst 33258 staining showed obvious morphological changes in U2-OS cells, including karyopyknosis and nuclear condensation. Flow cytometry showed a dose-dependent increase in the apoptosis rate of U2-OS cells treated with FP. Furthermore,the inhibitory effect of FP + DDP on the proliferation of U2-OS cells was time-dependent (P<C〇. 05). Moreover, the inhibitory rate,apoptosis rate and cycle arrest in FP + DDP group were significantly different from those in FP or DDP treatment group (P<C0. 05). Conclusion FP treatment can inhibit proliferation and induce apoptosis in human osteoblastic U2-OS cells. In addition,FP and DDP have a synergistic effect on the proliferation and apoptosis of U2-OS cells