半枝莲总黄酮对非小细胞肺癌细胞增殖及迁移的影响

Effects of total flavonoids of Scutellaria barbata D.Don on proliferation and migration of non-small cell lung cancer cells

目的观察半枝莲总黄酮对非小细胞肺癌细胞增殖、迁移及凋亡的影响并对其机制进行初步探索。方法体外培养非小细胞肺癌A549细胞,分为阴性对照组、实验组、阳性对照组。阴性对照组用1640培养液200μL处理细胞,实验组分别用1,5,10,20,40μg/mL半枝莲总黄酮处理细胞24 h,阳性对照组用200μL 40μg/mL芦丁溶液处理细胞。采用四甲基偶氮唑盐微量酶反应比色法(MTT)观察细胞增殖情况;通过流式细胞术检测细胞凋亡情况;蛋白质印迹法(Western Blot)检测凋亡抑制蛋白Survivin和促凋亡蛋白酶Caspase?3蛋白的表达;迁移划痕实验检测迁移的情况。结果MTT检测结果显示,阳性对照组的增殖抑制率为(57.234±0.533)%,实验组的增殖抑制率分别为(14.852±1.059)%、(23.486±1.366)%、(36.396±0.321)%、(46.179±1.334)%和(51.530±1.054)%,随浓度增加抑制率显著升高,呈浓度依赖(F=1 408.303,P<0.001);流式细胞术检测细胞凋亡率,阴性对照组(9.09±0.50)%、阳性对照组(67.27±1.48)%、实验组分别为(24.62±0.70)%、(34.67±0.46)%、(46.92±0.29)%、(55.67±0.57)%和(59.29±0.73)%,实验组的细胞凋亡率均高于阴性对照组(F=1 467.681,P<0.001);Western Blot结果显示,实验组Survivin蛋白表达均低于阴性对照组,Caspase?3蛋白表达均高于对照组(F=34.176,P<0.001;F=57.730,P<0.001);迁移划痕实验结果显示,半枝莲总黄酮抑制肺腺癌A549细胞迁移并呈现剂量依赖性。结论半枝莲总黄酮可抑制人肺腺癌A549细胞的增殖,可能与Survivin蛋白降低和Caspase?3蛋白表达量升高有关;另外,半枝莲总黄酮可抑制人肺腺癌A549细胞的迁移。

Objective To study the effects of total flavonoids of Scutellaria barbata D.Don on the proliferation,migration and apop?tosis of non?small cell lung cancer cells and to explore the mechanism.Methods Non?small cell lung cancer A549 cells were cul?tured in vitro and assigned into negative control group,experimental group and positive control group.The negative control group was treated with 200μL 1640 medium,the experimental group was treated with 1,5,10,20,40μg/mL total flavonoids of Scutellar?ia barbata D.Don for 24 h,and the positive control group was treated with 200μL 40μg/mL rutin solution.Cell proliferation was observed by MTT assay.Cell apoptosis was detected by flow cytometry.The protein expression levels of Survivin and Caspase?3 were detected by Western blot.Transfer scratch test was adopted to detect the migration situation.Results MTT results showed that the inhibition rates were(57.234±0.533)%in the positive control group,and(14.852±1.059)%,(23.486±1.366)%,(36.396±0.321)%,(46.179±1.334)%and(51.530±1.054)%,respectively in the experimental group.The inhibition rate increased significantly with the increase of concentration,which was concentration?dependent(F=1 408.303,P<0.001).Cell apoptosis rates were(9.09±0.50)%in the negative control group,(67.27±1.48)%in the positive control group,and(24.62±0.70)%,(34.67±0.46)%,(46.92±0.29)%,(55.67±0.57)%and(59.29±0.73)%respectively in the experimental group.Cell apoptosis rates in the experimental group were all higher than that in the negative control group(F=1 467.681,P<0.001).The protein expression of Survivin in the experi?mental group was lower than that in the negative control group,and the protein expression of Caspase?3 was higher than that in the negative control group(F=34.176,P<0.001;F=57.730,P<0.001).Additonally,total flavonoids of Scutellaria barbata D.Don could inhibit the migration of A549 cells in a dose?dependent manner.Conclusions The proliferation of A549 cells can be inhibited by total flavonoids of Scutellaria barbata D.Don,which may be related to the decrease of Survivin protein and the increased expression of Caspase?3.In addition,total flavonoids of Scutellaria barbata D.Don could inhibit the migration of lung adenocarcinoma A549 cells.