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目的研究二乙酰二脱水卫矛醇(DADAG)对鸟氨酸脱羧酶(ODC)活性的影响及其对人前列腺癌PC-3M细胞增殖的作用,并初步探讨其可能的分子机制。方法采用酶促反应法测定DADAG对PC-3M细胞中ODC活性的影响,用SRB法观察DADAG对PC-3M细胞增殖的抑制作用,流式细胞术分析DADAG对PC-3M细胞周期的改变,Western blot方法检测DADAG对PC-3M细胞周期相关蛋白Cdc2,CyclinB1,p-Cdc2(Tyr15),Cdc25C和Wee1表达的影响。结果5和10mg·L-1DADAG作用PC-3M细胞24h后,ODC活性的抑制率分别为39.3%和31.1%。SRB结果显示,40mg·L-1DADAG作用PC-3M细胞48和72h后,细胞增殖抑制率分别为34.6%和47.8%,与对照组比较有显著性差异(P<0.05)。细胞周期分析表明,5,10和20mg·L-1DADAG作用48h后,G2/M期细胞百分数为47.4%,62.9%和69.1%。DADAG作用细胞后,Cdc25C的表达下调,而Cyclin B1,p-Cdc2(Tyr15)和Wee1表达上调,Cdc2变化不明显。结论DADAG可降低PC-3M细胞中ODC的活性,并抑制细胞的增殖,使阻滞细胞周期于G2/M期,DADAG的抗肿瘤机制与降低ODC活性,改变p-Cdc2(Tyr15),Cdc25C和Wee1蛋白表达有关。
Objective To investigate the effect of diacetyldianhydrogalactitol (DADAG) on ornithine decarboxylase (ODC) activity and its effect on the proliferation of human prostate cancer PC-3M cells and to explore its possible molecular mechanism. Methods The effect of DADAG on the activity of ODC in PC-3M cells was determined by enzymatic reaction. The inhibitory effect of DADAG on the proliferation of PC-3M cells was observed by SRB method. The cell cycle of DADAG-treated PC-3M cells was analyzed by flow cytometry. blot was used to detect the effect of DADAG on the expression of cyclin B1, p-Cdc2 (Tyr15), Cdc25C and Wee1 in PC-3M cells. Results The inhibitory rates of ODC activity of PC-3M cells treated with 5 and 10 mg · L-1 DADAG for 24 h were 39.3% and 31.1%, respectively. The results of SRB showed that the cell proliferation inhibition rates of PC-3M cells treated with 40 mg · L-1 DADAG for 48 and 72 h were 34.6% and 47.8%, respectively, which were significantly different from the control group (P <0.05). Cell cycle analysis showed that the percentages of cells in G2 / M phase were 47.4%, 62.9% and 69.1% after treated with 5, 10 and 20 mg · L-1 DADAG for 48 h. After DADAG treatment, the expression of Cdc25C was down-regulated while the expressions of Cyclin B1, p-Cdc2 (Tyr15) and Wee1 were up-regulated while the changes of Cdc2 were not obvious. Conclusion DADAG can reduce the activity of ODC in PC-3M cells and inhibit the proliferation of cells. It can block the cell cycle in G2 / M phase, inhibit the anti-tumor mechanism of DADAG and decrease the activity of ODC, and change the expression of p-Cdc2 (Tyr15), Cdc25C and Wee1 protein expression.