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目的探讨丹皮酚对人卵巢癌A2780细胞的生长抑制及诱导凋亡作用,揭示Wnt/β-catenin信号转导通路分子机制。方法对数生长期人卵巢癌A2780细胞分为对照组(0mmol/L丹皮酚)和0.4、0.8、1.6mmol/L丹皮酚组,采用CCK-8法检测丹皮酚作用24、48、72h时人卵巢癌A2780细胞增殖活性;丹皮酚作用48h后,应用流式细胞仪检测人卵巢癌A2780细胞周期,采用Western blot法检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2associated X protein,Bax)、β-catenin和c-Myc蛋白表达水平;丹皮酚作用0、24、48h,采用划痕实验检测人卵巢癌A2780细胞迁移能力。结果丹皮酚作用24、48、72h,人卵巢癌A2780细胞增殖活性在对照组为(100.00±3.28)%、(100.00±2.34)%、(100.00±2.89)%,在0.4 mmol/L丹皮酚组为(93.26±4.21)%、(80.23±5.43)%、(72.04±4.89)%,在0.8mmol/L丹皮酚组为(81.25±3.13)%、(71.65±4.78)%、(62.32±4.39)%,在1.6mmol/L丹皮酚组为(60.85±2.35)%、(45.89±5.34)%、(35.74±3.19)%,除0.4mmol/L丹皮酚组作用24h外,0.4、0.8、1.6mmol/L丹皮酚组不同作用时间人卵巢癌A2780细胞增殖活性均低于对照组(P<0.05),0.8 mmol/L丹皮酚组作用24h,1.6mmol/L丹皮酚组作用24、48、72h人卵巢癌A2780细胞增殖活性均低于0.4 mmol/L丹皮酚组(P<0.05),1.6mmol/L丹皮酚组作用24、48、72h人卵巢癌A2780细胞增殖活性均低于0.8 mmol/L丹皮酚组(P<0.05),0.4、0.8、1.6mmol/L丹皮酚组作用72h人卵巢癌A2780细胞增殖活性低于24h(P<0.05);丹皮酚作用48h,0.4、0.8、1.6mmol/L组G1期细胞比例(73.08%,68.46%、52.95%)低于对照组(78.30%),S期细胞比例(19.61%、26.11%、39.05%)高于对照组(17.48%)(P<0.05);丹皮酚作用48h,0.4、0.8、1.6mmol/L组Bax蛋白表达明显高于对照组,0.8、1.6mmol/L组Bcl-2、β-catenin和c-Myc蛋白表达明显低于对照组(P<0.05);划痕实验结果表明,细胞迁移能力随丹皮酚浓度增加和作用时间延长明显下降(P<0.05)。结论丹皮酚可抑制人卵巢癌A2780细胞增殖并促进其凋亡,其机制可能与抑制Wnt/β-catenin信号转导通路有关。
Objective To investigate the inhibitory effect of paeonol on the growth of human ovarian cancer A2780 cells and to reveal the molecular mechanism of Wnt / β-catenin signal transduction pathway. Methods A2780 cells in logarithmic growth phase were divided into control group (0mmol / L paeonol) and 0.4,0.8,1.6mmol / L paeonol group. The effect of paeonol was determined by CCK-8 assay 24,48, 72h human ovarian cancer A2780 cell proliferation; paeonol after 48h, flow cytometry was used to detect cell cycle of human ovarian cancer A2780 cell cycle, B-cell lymphoma-2 (Bcl-2, Bcl- -2), Bcl-2associated X protein (Bax), β-catenin and c-Myc protein expression levels; paeonol 0,24,48h, scratch test was used to detect human ovarian cancer A2780 Cell migration ability. Results The proliferation activity of paeonol at 24,48,72 h and the proliferation of human ovarian cancer cell line A2780 in the control group were (100.00 ± 3.28)%, (100.00 ± 2.34)% and (100.00 ± 2.89)%, respectively. (81.25 ± 3.13)%, (71.65 ± 4.78)%, (62.32 ± 0.83)% in the group of 0.8 mmol / 4.39%, respectively. Compared with 0.4mmol / L paeonol group, the activity of paeonol in 0.4mmol / L paeonol group was (0.4) The proliferation activities of A2780 cells in 0.8 and 1.6 mmol / L paeonol groups for different time were lower than those in the control group (P <0.05), paeonol group in 0.8 mmol / L paeonol group for 24h and 1.6 mmol / L paeonol group Proliferation activity of A2780 cells in 24, 48 and 72h human ovarian cancer cells was lower than that in 0.4 mmol / L paeonol group (P <0.05) (P <0.05). The proliferation activity of A2780 cells treated with 0.4, 0.8 and 1.6 mmol / L paeonol for 72 h was lower than that of 24h (P <0.05) The percentage of cells in G1 phase (73.08%, 68.46%, 52.95%) in phenol group was lower than that in control group (78.30%) in 48h, 0.4,0.8 and 1.6mmol / (P <0.05). The expression of Bax in paeonol treated group was significantly higher than that in control group at 48 h, 0.4, 0.8 and 1.6 mmol / L, The expression of Bcl-2, β-catenin and c-Myc protein in 1.6 mmol / L group was significantly lower than that in control group (P <0.05). Scratch test results showed that cell migration ability decreased significantly with the increase of paeonol concentration and the prolongation of action time (P <0.05). Conclusion Paeonol can inhibit the proliferation and promote the apoptosis of human ovarian cancer cell line A2780, which may be related to the inhibition of Wnt / β-catenin signal transduction pathway.