论文部分内容阅读
目的探讨EZH2抑制剂DZNe P对人乳腺癌细胞增殖、凋亡及相关信号通路的影响。方法取对数生长期乳腺癌细胞MCF-7、MDA-MB-231,经2.5、5、10、20μmol/L DZNe P处理后(设不加DZNe P仅添加完全培养基组为对照组),采用四甲基偶氮唑盐比色法检测各组24、48、72 h的增殖抑制率,Annexin V-FITC/PI流式细胞术检测各组24、48 h的凋亡率,收集20μmol/L DZNe P处理72 h的MCF-7、MDA-MB-231细胞,采用Western印迹检测磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路和Wnt/β-连接素(β-catenin)通路中的蛋白变化。结果 DZNe P在2.5~20μmol/L范围内可抑制MCF-7和MDA-MB-231细胞增殖,且与对照组相比增殖抑制率升高(P<0.05),该抑制效应呈浓度和时间依赖性;与对照组相比,DZNe P处理组的凋亡率均升高(P<0.05),DZNe P各浓度处理48 h的凋亡率均高于24 h(P<0.05);PI3K/Akt通路中,与对照组相比,20μmol/L DZNe P处理72 h后MCF-7、MDA-MB-231细胞的PTEN水平均升高,Akt水平均降低(P<0.05);而在Wnt/β-catenin通路上,20μmol/L DZNe P处理72 h后两细胞株中的β-catenin、Cyclin D1和C-myc水平均降低(P<0.05)。结论 DZNe P可抑制人乳腺癌细胞增殖并诱导其凋亡,可能与抑制肿瘤恶性行为相关的PI3K/Akt、Wnt/β-catenin通路有关,在乳腺癌治疗上有一定应用前景。
Objective To investigate the effects of EZH2 inhibitor DZNe P on proliferation, apoptosis and related signal pathways in human breast cancer cells. Methods After treated with 2.5, 5, 10, 20μmol / L DZNe P, MCF-7 and MDA-MB-231 cells in logarithmic growth phase were treated with DZNe P supplemented with complete medium The proliferation inhibition rate of each group was detected by MTT colorimetric assay at 24, 48, and 72 h. The apoptosis rate of 24, 48, and 48 h in each group was detected by Annexin V-FITC / PI flow cytometry. L-DZNe P for 72 h. The expression of PI3K / Akt pathway and Wnt / β-catenin (β-catenin) ) Pathway in the protein changes. Results DZNe P could inhibit the proliferation of MCF-7 and MDA-MB-231 cells in the range of 2.5 ~ 20μmol / L, and the inhibitory rate of proliferation increased compared with the control group (P <0.05). The inhibitory effect was dose- and time-dependent (P <0.05). Compared with the control group, the apoptosis rate of DZNe P group was higher than that of the control group (P <0.05) Pathway, compared with the control group, PTEN levels in MCF-7 and MDA-MB-231 cells increased and Akt levels decreased at 20 μmol / L DZNe P treatment for 72 h (P <0.05) -catenin pathway, the levels of β-catenin, Cyclin D1 and C-myc in both cell lines decreased after treatment with 20 μmol / L DZNe P for 72 h (P <0.05). Conclusion DZNe P can inhibit the proliferation and induce the apoptosis of human breast cancer cells, which may be related to the inhibition of the PI3K / Akt and Wnt / β-catenin pathways that may be involved in the malignant behavior of breast cancer. Therefore, DZNe P has some potential applications in the treatment of breast cancer.