雷公藤内酯醇对小鼠前列腺癌细胞株RM-1的影响

目的:探讨雷公藤内酯醇(TL)对前列腺癌细胞株RM-1增殖的影响,分析其抗前列腺癌细胞的作用机制。方法:MTT实验检测TL对RM-1细胞的增殖抑制;吖啶橙染色检测细胞形态学变化;FCM分析细胞周期及凋亡峰;DNA电泳分析细胞凋亡断裂带;RT-PCR检测RM-1细胞中caspase-3及bcl-2 mRNA表达。结果:MTT实验结果显示,TL(5、10、20、40、80ng/ml)处理后RM-1细胞生长抑制率分别为9.8%、25.1%、39.2%、48.8%、53.2%,以TL(10、20ng/ml)处理RM-1细胞12、24、36、48h后,细胞生长抑制率分别为8.4%、25.1%、36.1%、42.4%和10.2%、39.2%、50.2%、58.5%;经TL处理后吖啶橙染色观察发现,细胞核浓缩,细胞膜内陷、胞内有不规则橘黄色颗粒,呈凋亡样形态学改变;FCM检测显示TL(10、20ng/ml)作用RM-1细胞48h后,在G1期前出现明显的凋亡峰;TL处理RM-1细胞24、36、48h后,均可见DNA“梯形”条带;TL处理RM-1细胞caspase-3 mRNA表达高于未处理对照细胞(P<0.05),bcl-2 mRNA表达低于对照细胞(P<0.05)。结论:TL可抑制小鼠前列腺癌细胞株RM-1增殖,此作用是通过降低bcl-2、提高caspase-3表达诱导小鼠前列腺癌细胞凋亡来实现的。

Effects of Triptolide on Prostate Carcinoma in Mouse RM-1 Cells

OBJECTIVE: To study the effect of Triptolide (TL) on the growth of prostate carcinoma cell line, and analyze its function and mechanism in anti-prostate cancer. METHODS: MTT experiments were performed to examine the inhibiting effect of TL on the proliferation of RM-1 cells, cell morphological changes observed by acridine staining, cellular cycles and apoptosis peak analyzed by flow cytometry, the apoptosis fracture zone investigated with DNA electrophoresis, and the expressions of caspase-3 and bcl-2 mRNA in RM-1 cells examined by RT-PCR. RESULTS: The results of MTT experiments showed that after the treatment of TL (5, 10, 20, 40 and 80 ng/ml), the RM-1 cell proliferation inhibition rates were 9.8%, 25.1%, 39.2%, 48.8% and 53.2% respectively; 12, 24, 36 and 48 hours after the treatment of TL (10 and 20 ng/ml), the cell proliferation inhibition rates were 8.4%, 25.1%, 36.1%, 42.4% and 10.2%, 39.2%, 50.2% and 58.5% respectively. Acridine staining after the TL treatment revealed nucleus condensation, cell membrane invagination, irregular orange particles in the cells and apoptosis morphological changes; flow cytometry tests showed that 48 hours after the TL treatment (10, 20 ng/ml) of RM-1 cells, an obvious apoptosis peak appeared before the G1 stage; 24, 36 and 48 hours after it, DNA "trapezoid" strips could be seen; the caspase-3 mRNA expression in the TL treated cells was higher, and the bcl-2 mRNA expression was lower than in the controls. CONCLUSION: TL can decrease bcl-2 expression, increase caspase-3 expression, induce apoptosis of prostate carcinoma cells, and consequently inhibit the proliferation of RM-1 cells in mice.