毛蕊异黄酮对肺腺癌SPC-A1细胞增殖和凋亡的影响

Effect of calycosin on proliferation and apoptosis of lungadenocarcinoma cell line SPC-A1

目的探讨毛蕊异黄酮对肺腺癌SPC-A1细胞增殖与凋亡的影响。方法将25、50及100μg/ml毛蕊异黄酮分别作用SPC-A1细胞12、24、36及48 h后,用MTT法检测其对SPC-A1细胞增殖的抑制作用;Hoechst 33258染色观察SPC-A1细胞核变化,流式细胞术检测SPC-A1细胞凋亡情况;不同浓度毛蕊异黄酮作用SPC-A1细胞48 h后,Western blotting及逆转录聚合酶链反应(RT-PCR)检测SPC-A1细胞Cyclin D1、CDK4、CDK6蛋白及mRNA的表达水平。结果毛蕊异黄酮能有效抑制SPC-A1细胞的增殖,且呈浓度和时间依赖性(P<0.05),100μg/ml毛蕊异黄酮作用SPC-A1细胞48 h后,细胞增殖抑制率可高达(81.26±0.05)%;毛蕊异黄酮能够诱导SPC-A1细胞凋亡,引起核固缩,形成凋亡小体,细胞凋亡率从(3.12±1.07)%升至(42.78±2.16)%(P<0.05);毛蕊异黄酮可以抑制SPC-A1细胞中Cyclin D1、CDK4、CDK6蛋白及mRNA的表达,组间比较差异有统计学意义(P<0.05)。结论毛蕊异黄酮可通过降低Cyclin D1、CDK4、CDK6蛋白及mRNA的表达抑制SPC-A1细胞的增殖,毛蕊异黄酮对SPC-A1细胞具有诱导凋亡作用。

Objective To investigate the effect of calycosin on proliferation and apoptosis of lung adenocarcinoma cells line SPC-A1 in vitro.Methods After treatment with different concentrations of calycosin(25,50 and 100μg/ml)for different time(12,24,36 and 48 h),the proliferation of lung adenocarcinoma SPC-A1cells were detected by MMT assay;Hoechst33258 fluorescent staining was employed to observe the nucleus;the cells apoptosis were detected by flow cytometry;Western blot and real-time PCR experiment were used to analyze the expression of calycosin levels of Cyclin D1,CDK4,CDK6 protein and mRNA in SPC-A1 cells with(25,50 and 100μg/ml)calycosin for 48 h.Results MTT experiment showed that the cell proliferation of SPC-A1 cells was inhibited by calycosin,and in a dose-dependent and time-dependent manner(P<0.05),with inhibitory rate of(81.26±0.05)%at 100μg/ml calycosin for 48 h;calycosin could induce apoptosis of SPC-A1 cells,cause the nucleus pycnosis,produce apoptosis bodies,and the apoptosis rate of SPC-A1 cells increased from(3.12±1.07)%to(42.78±2.16)%(P<0.05)as detected by flow cytometry;Western blotting and RT-PCR experiment demonstrated that the expression of Cyclin D1,CDK4,CDK6 of SPC-A1 cells can be inhibited by calycosin,with significant difference among groups(P<0.05).【Conclusions】Calycosin can inhibit the proliferation of SPC-A1 cells by inhibiting expression of Cyclin D1,CDK4,CDK6 protein and their mRNA,and induce apoptosis of SPC-A1 cells.