论文部分内容阅读
目的:观察Akt抑制剂鱼藤素对前列腺癌PC-3细胞株的抑制作用并探讨其可能的作用机制。方法:用MTT法检测鱼藤素对PC-3细胞的增殖抑制率;流式细胞术(FCM)检测鱼藤素对细胞周期的影响;RT-PCR检测细胞中小鼠双微体2(MDM2)、糖元合成酶激酶3β(GSK3β)mRNA表达的变化;Western印迹法检测MDM2、GSK3β蛋白表达的变化。结果:MTT法显示,10、100、500、1 000 nmol/L的鱼藤素作用于前列腺癌PC-3细胞24、48、72 h后,对前列腺癌PC-3细胞增殖抑制率分别为(91.10±3.75)%、(86.39±1.16)%、(79.51±2.63)%;(82.46±3.65)%、(76.84±0.97)%、(69.69±2.30)%;(81.46±0.41)%、(75.56±1.12)%、(54.07±3.21)%;(66.77±2.82)%、(58.22±0.35)%、(39.34±2.40)%,均能显著抑制其增殖(P均<0.01);FCM检测显示各浓度的鱼藤素使前列腺癌PC-3细胞周期阻滞在G0/G1期比例增加[(53.4±2.3)%、(62.4±2.2)%、(63.6±1.1)%、(65.0±0.3)%、(66.5±1.9)%,P均<0.01],S期细胞比例减少[(26.9±1.7)%、(14.7±2.4)%、(11.1±5.2)%、(5.8±1.1)%、(7.0±0.6)%,P均<0.01];RT-PCR和Western印迹法结果显示鱼藤素上调了GSK3βmRNA和蛋白的表达水平,而下调了MDM2 mRNA和蛋白的表达水平。结论:Akt抑制剂鱼藤素能抑制前列腺癌PC-3细胞的增殖,其机制可能与影响Akt信号通路下游分子GSK3β、MDM2的表达相关。
Objective: To observe the inhibitory effect of Akt inhibitor deguelin on prostate cancer cell line PC-3 and to explore its possible mechanism. Methods: The inhibitory rate of deguelin on proliferation of PC-3 cells was determined by MTT assay. The effect of deguelin on cell cycle was detected by flow cytometry (FCM). The expression of MDM2 in mouse cells was detected by RT- , Glycogen synthase kinase 3β (GSK3β) mRNA expression were detected by Western blotting to detect the expression of MDM2, GSK3βprotein. Results: MTT assay showed that the inhibitory rates of 10, 100, 500, 1000 nmol / L deguelin on prostate cancer PC-3 cells were 24, 48, 72 h after prostatic carcinoma PC-3 cells proliferation respectively 91.8 ± 3.75%, (86.39 ± 1.16)%, (79.51 ± 2.63)%; (82.46 ± 3.65)%, (76.84 ± 0.97)%, (69.69 ± 2.30)%, (81.46 ± 0.41)%, (P <0.01). The FCM results showed that each of the three groups had significant difference (P <0.01) (53.4 ± 2.3)%, (62.4 ± 2.2)%, (63.6 ± 1.1)%, (65.0 ± 0.3)% respectively in the G0 / G1 phase of PC- (66.5 ± 1.9)%, P <0.01]. The percentage of cells in S phase decreased (26.9 ± 1.7)%, (14.7 ± 2.4)%, (11.1 ± 5.2)%, ± 0.6%, P <0.01]. RT-PCR and Western blotting showed that deguelin upregulated the expression of GSK3β mRNA and protein, but down-regulated the expression of MDM2 mRNA and protein. Conclusions: The inhibition of Akt inhibitor deguelin can inhibit the proliferation of PC-3 cells. The mechanism may be related to the expression of GSK3β and MDM2.