土槿乙酸抑制人卵巢癌SKOV_3细胞增殖并诱导G_2/M期阻滞

目的探讨土槿乙酸(pseudolaric acid B,PLAB)对人卵巢癌SKOV3细胞增殖和细胞周期的影响。方法 MTT法检测PLAB对细胞生长抑制作用;Hoechst 33342荧光染色分析细胞死亡特征;流式细胞术检测细胞周期;Real time-PCR和Western blot检测周期相关基因Cyclin B1、CDK1和CyclinD1的表达变化。结果 PLAB明显抑制SKOV3细胞生长,且抑制作用呈浓度-作用时间依赖性(P<0.05),其24、48和72 h的IC50值分别为2.44、1.60、2.94μmol.L-1。5μmol.L-1PLAB处理细胞24 h,细胞出现核染色质凝集、凋亡小体等典型凋亡特征,随着PLAB浓度升高,G2/M期细胞比例明显增大并表现出时间依赖性(P<0.05),同时伴有Cyclin B1和CDK1呈高表达状态(P<0.05),Cyclin D1呈低表达状态(P<0.05)。结论 PLAB可抑制SKOV3细胞生长,引起细胞G2/M期阻滞,伴随Cyclin B1和CDK1呈高表达状态,可能与其调控周期相关蛋白降解有关。

Growth inhibition and G2/M phase arrest in human ovarian cancer SKOV3 cells induced by pseudolaric acid B

Aim To study the cell cycle change of human ovarian cancer SKOV3 cells treated with pseudolaric acid B(PLAB),and to explore its mechanisms.Methods The cell growth inhibition was measured by MTT method.Cell morphologic changes were determined by Hoechest33342 staining.The cell cycle was measured by flow cytometry.The mRNA and protein expression levels of Cyclin B1,Cyclin D1 and CDK1 were detected by Real time PCR and Western blot respectively.Results PLAB could inhibit SKOV3 cell growth in a dose-and time-dependent manner(P<0.05).The IC50 values of PLAB for SKOV3 cells for 24,48,72 h were 2.44,1.60,2.94 μmol·L-1,respectively.SKOV3 cell treated with 5 μmol·L-1 PLAB for 24 h showed some typical apoptosis characteristics such as nuclei chromatin condensation and apoptotic body.The proportion of G2/M phase cells strikingly increased in a dose-and time dependent manner(P<0.05),meanwhile the Cyclin B1 and CDK1 expressions exhibited higher levels(P<0.05),and the Cyclin D1 expression exhibited lower levels(P<0.05).Conclusion PLAB can inhibit the proliferation of SKOV3 cells and generate the cell cycle G2/M arrest,which may be related with its regulation of cell cycle-associated proteins.