香附提取物在表柔比星促进三阴性乳腺癌细胞凋亡中的作用及机制  

Effect of Cyperus Rotundus L. extract on epirubicin in promoting apoptosis of triple negative breast cancer cells

作　　者：边梦雪 于志勇 BIAN Mengxue;YU Zhiyong(School of Medicine and Life Sciences,University of Jinan,Jinan 250017,China)

机构地区：[1]济南大学医学与生命科学学院,济南250117 [2]山东省医学科学院 [3]山东大学附属山东省肿瘤医院

出　　处：《山东医药》2020年第1期44-47,共4页Shandong Medical Journal

基　　金：山东省自然科学基金资助项目(ZR2019MH109)

摘　　要：目的探讨香附提取物在表柔比星促进三阴性乳腺癌细胞凋亡中的作用及机制。方法常规培养三阴性乳腺癌细胞株MDA-MB-231,分别加入剂量为0、0.2、0.4、0.6、0.8、1.2 mg/mL香附提取物,0、0.75、1.5、3μg/mL表柔比组,获得香附提取物、表柔比星接近半数致死量分别为0.4 mg/mL、1.2μg/mL。取部分细胞用0.4 mg/mL香附+1.2μg/mL表柔比星处理48 h。用CCK-8实验检测细胞增殖活力,用流式细胞术检测细胞凋亡率,用细胞平板克隆形成实验检测细胞克隆形成率;Western blotting法检测细胞内凋亡标志蛋白Bax、抗凋亡标志蛋白Bcl-2及自噬标志蛋白Beclin-1表达。结果香附提取物联合表柔比星处理后,MDA-MB-231细胞存活率低于二者单独处理(P均<0.05)。香附提取物细胞半数致死量为0.4 mg/mL、表柔比星半数致死量为1.25μg/mL。0.4 mg/mL香附提取物+1.2μg/mL表柔比星处理后,MDA-MB-231细胞早期凋亡率、晚期凋亡率、总凋亡率均高于二者单独处理(P均<0.05)。经0.4 mg/mL香附提取物、1.2μg/mL表柔比星、0.4 mg/mL香附提取物+1.2μg/mL表柔比星处理后,MDA-MB-231克隆形成率分别为(65.32±6.41)%、(69.52±5.41)%、(45.68±8.31)%。经0.4 mg/mL香附提取物+1.2μg/mL表柔比星处理后MDA-MB-231克隆形成率低于二者单独处理(P均<0.05)。经0.4 mg/mL香附提取物+1.2μg/mL表柔比星处理后MDA-MB-231细胞内Bax蛋白表达高于二者单独处理,Bcl-2、Beclin-1蛋白表达低于二者单独处理(P均<0.05)。结论香附提取物可增强表柔比星对三阴性乳腺癌细胞的促凋亡作用,其机制可能与调控凋亡相关蛋白表达及抵制细胞自噬有关。Objective To investigate the effect and mechanism of Cyperus Rotundus L.extract(Cyperus extract)on the apoptosis of triple negative breast cancer cells.Methods Triple negative breast cancer cells MDA-MB-231 were cultured in a conventional manner.The cells were treated with 0,0.2,0.4,0.6,0.8 and 1.2 mg/mL Cyperus extract,0,0.75,1.5 and 3μg/mL epirubicin,and 0.4 mg/mL Cyperus extract combined with epirubicin 1.2μg/mL for 48 h,respectively.CCK-8 assay was used to detect cell proliferation activity,flow cytometry was used to detect apoptosis,cell plate clone formation assay was used to detect cell clone formation,and Western blotting was used to detect the expression of Bax,Bcl-2 and Beclin-1.Results The survival rate of MDA-MB-231 cells treated with Cyperus extract and epirubicin was lower than that of the two treatments alone(P<0.05).The inhibitory concentration 50(IC 50)of Cyperus extract on cells was 0.4 mg/mL and that of epirubicin was 1.25μg/mL.After treatment with 0.4 mg/mL Cyperus extract and 1.2μg/mL epirubicin,the early apoptosis rate,late apoptosis rate and total apoptosis rate of MDA-MB-231 cells were higher than those of MDA-MB-231 cells treated alone(P<0.05).The clone formation rate of MDA-MB-231 were 65.32%±6.41%,69.52%±5.41%,45.68%±8.31%,respectively.The clone formation rate of MDA-MB-231 treated with 0.4 mg/ml Cyperus extract and 1.2μg/ml epirubicin was lower than that of the two treatments alone(P<0.05).The expression of Bax in MDA-MB-231 cells treated with 0.4 mg/mL extract of Cyperus and 1.2μg/mL epirubicin was higher than that of the two treatments alone,as well as the expression of Bcl-2 and Beclin-1(all P<0.05).Conclusion Cyperus extract can enhance the apoptotic effect of epirubicin on three negative breast cancer cells,which may be related to the regulation of apoptosis-related protein expression and autophagy.

关 键 词：三阴性乳腺癌 香附提取物 表柔比星 细胞凋亡 细胞自噬 

分 类 号：R737.9[医药卫生—肿瘤]