β-紫罗兰酮通过Caspase-3信号通路对乳腺癌细胞生物学行为的调控作用

Analysis of Biological Behaviors of Human Breast Cancer Cell Line Under the Regulation of β-ionone Via Caspase-3 Signaling Pathway

目的研究β-紫罗兰酮通过Caspase-3信号对乳腺癌细胞生物学行为的调控作用。方法将人乳腺癌MCF-7细胞分为空白对照组与实验组,实验组按照β-紫罗兰酮的不同浓度,又分为25μmol/L组、50μmol/L组、100μmol/L组及200μmol/L组。分别采用CCK-8及台盼蓝计数法检测β-紫罗兰酮对MCF-7细胞增殖及生长曲线的影响,Hoechst 33258荧光染色法观察β-紫罗兰酮对MCF-7细胞凋亡形态的影响,RT-PCR及Western Blot法检测β-紫罗兰酮对Caspase-3 mRNA及蛋白表达的影响。结果与空白对照组相比,经不同浓度β-紫罗兰酮处理后的乳腺癌MCF-7细胞OD值均显著下降(P<0.05),实验组中经β-紫罗兰酮浓度25μmol/L、50μmol/L、100μmol/L及200μmol/L处理后的MCF-7细胞增殖抑制率分别为7.92%、28.96%、45.22%和56.83%。不同浓度β-紫罗兰酮处理过的MCF-7细胞从第2天开始,较空白对照组的细胞计数显著降低(P<0.05),空白组对照组及β-紫罗兰酮浓度为25μmol/L组MCF-7细胞计数在7 d内随时间迁移呈上升趋势,经β-紫罗兰酮浓度为50、100及200μmol/L处理后的MCF-7细胞7 d内细胞计数呈下降趋势。Hoechst 33258荧光染色法检测发现,与空白对照组相比,给予β-紫罗兰酮处理过的人乳腺癌MCF-7细胞伴有不同程度的凋亡现象,而经β-紫罗兰酮浓度为200μmol/L处理后的细胞凋亡现象最明显。与空白对照组相比,经不同浓度β-紫罗兰酮处理过的MCF-7细胞Caspase-3 mRNA及蛋白表达水平均显著上升(P<0.05),且随β-紫罗兰酮浓度的升高,其Caspase-3 mRNA及蛋白表达呈上升趋势(P<0.05)。结论β-紫罗兰酮可能通过激活Caspase-3信号通路,抑制乳腺癌细胞增殖与生长,促进其凋亡,发挥抑癌作用。

Objective To investigate the regulation ofβ-ionone on regulating the biological behavior of breast cancer cells through caspase-3 signaling.Methods Human breast cancer MCF-7 cells were divided into blank control group and the experimental group.The experimental group was further divided into 4 subgroups according to the different concentrations ofβ-ionone,25μmol/L,50μmol/L,100μmol/L and 200μmol/L subgroups.CCK-8 and trypan blue exclusion methods were used to detect the effects ofβ-ionone on the proliferation and growth curve of MCF-7 cells.Hoechst 33258 fluorescent staining was used to observe the effects ofβ-ionone on the apoptotic morphology of MCF-7 cells.Meantime,RT-PCR and Western Blot were used to detect the effect ofβ-ionone on the mRNA and protein expressions of Caspase-3.Results Compared with blank control group,the optical density(OD)values of breast cancer MCF-7 cells treated with different concentrations ofβ-ionone was significantly decreased(P<0.05).The proliferation inhibition rates of MCF-7 cells were 7.92%,28.96%,45.22%and 56.83%under the treatment withβ-ionone at the concentrations of 25μmol/L,50μmol/L,100μmol/L and 200μmol/L in experimental groups,respectively.The cell counts of MCF-7 cells treated with different concentrations ofβ-ionone was significantly lower than that of blank control group from the second day of treatment(P<0.05).During the 7 days of treatment,MCF-7 cell counts in blank control group and 25μmol/L subgroup showed an increasing trend over time,while MCF-7 cell counts in 50μmol/L,100μmol/L and 200μmol/L subgroups showed a decreasing trend.Hoechst 33258 fluorescence staining showed that human breast cancer MCF-7 cells treated withβ-ionone showed varying degrees of apoptosis compared to blank control group,with the most obvious apoptosis occurring in cells treated withβ-ionone at a concentration of 200μmol/L.The mRNA and protein expression levels of Caspase-3 in MCF-7 cells treated with different concentrations ofβ-ionone increased significantly compared to blank control group(P<0.05),and the increase was in a concentration-dependent manner(P<0.05).Conclusionβ-ionone may inhibit the proliferation and growth of breast cancer cells and promote their apoptosis by activating the Caspase-3 signaling pathway,thereby exerting a tumor suppressor effect.