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目的:观察槲皮素对人前列腺癌PC-3细胞增殖和Wnt/β-连环蛋白(β-catenin)信号通路的调控作用,探讨其抗前列腺癌的可能机制。方法:体外培养PC-3细胞,以不同终浓度槲皮素进行处理,设空白组。采用噻唑蓝比色法(MTT)观察槲皮素对PC-3细胞增殖抑制作用,计算半数抑制浓度(IC50);采用流式细胞仪检测槲皮素对PC-3细胞凋亡的影响;采用免疫印迹法(Western blot)和逆转录-PCR(RT-PCR)分析检测槲皮素对β-catenin和下游靶基因细胞周期素D1(Cyclin D1),原癌基因(c-Myc)蛋白表达和转录水平;采用免疫荧光分析检测槲皮素对β-catenin表达的影响;利用荧光酶报告基因分析检测槲皮素对Wnt/β-catenin信号通路的影响。结果:与空白组比较,槲皮素对PC-3细胞增殖具有明显抑制作用(P<0.05),并诱导PC-3细胞凋亡(P<0.05)。与空白组比较,槲皮素可显著抑制PC-3细胞β-catenin的蛋白表达和转录活性,从而阻断Wnt/β-catenin信号通路的传导,抑制下游靶基因的表达水平(P<0.05,P<0.01)。结论:槲皮素可显著抑制PC-3细胞增殖,其机制可能与抑制Wnt/β-catenin信号通路有关。
AIM: To investigate the regulatory effect of quercetin on proliferation and Wnt / β-catenin signaling pathway in human prostate cancer PC-3 cells and to explore its possible mechanism of anti-prostate cancer. Methods: PC-3 cells were cultured in vitro and treated with different concentrations of quercetin. A blank group was established. The inhibitory effect of quercetin on the proliferation of PC-3 cells was observed by MTT and the IC50 was calculated. The effect of quercetin on the apoptosis of PC-3 cells was detected by flow cytometry. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of cyclin D1 and c-Myc protein and the protein level of β-catenin and downstream target genes The effect of quercetin on the expression of β-catenin was detected by immunofluorescence assay. The effect of quercetin on the Wnt / β-catenin signaling pathway was detected by luciferase reporter assay. Results: Compared with the blank group, quercetin significantly inhibited the proliferation of PC-3 cells (P <0.05) and induced the apoptosis of PC-3 cells (P <0.05). Compared with the blank group, quercetin significantly inhibited the protein expression and transcriptional activity of β-catenin in PC-3 cells, thereby blocking the Wnt / β-catenin signaling pathway and inhibiting the expression of downstream target genes (P <0.05, P <0.01). Conclusion: Quercetin can significantly inhibit the proliferation of PC-3 cells, which may be related to the inhibition of Wnt / β-catenin signaling pathway.