芹菜素与曲妥珠单抗联用对HER2阳性乳腺癌协同抗肿瘤作用  

Synergistic antitumor effect of apigenin combined with trastuzumab on HER2-positive breast cancer

作　　者：张梦瑶 雷碧黠 解伟 梁蓓蓓[2] 李博华 ZHANG Mengyao;LEI Bixia;XIE Wei;LIANG Beibei;LI Bohua(Graduate School,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Key Laboratory for Molecular Imaging Shanghai University of Medicine&Health Sciences,Shanghai 201318,China)

机构地区：[1]上海中医药大学研究生院,上海201203 [2]上海健康医学院分子影像学重点实验室,上海201318

出　　处：《中华肿瘤防治杂志》2024年第7期405-413,共9页Chinese Journal of Cancer Prevention and Treatment

基　　金：上海市卫健委科研项目(20214Y0516);上海市教委“晨光计划”(18-CG72)。

摘　　要：目的探讨芹菜素与曲妥珠单抗联合应用对人表皮生长因子受体2(HER2)阳性乳腺癌细胞系BT474及SK-BR-3的抗肿瘤作用及其机制。方法采用流式细胞术检测曲妥珠单抗与人乳腺癌细胞的结合活性;细胞计数试剂盒(CCK-8)检测芹菜素、曲妥珠单抗单药及联合应用对BT474和SK-BR-3细胞的生长抑制活性;EdU染色法检测芹菜素、曲妥珠单抗单药及联合应用对BT474和SK-BR-3细胞DNA复制的影响;蛋白质印迹法检测芹菜素、曲妥珠单抗单药及联合应用对BT474和SK-BR-3细胞中表皮生长因子受体(EGFR)、HER2、HER3、Akt等蛋白及其各自磷酸化蛋白表达水平的影响。设置芹菜素组、曲妥珠单抗组、芹菜素+曲妥珠单抗联合用药组、对照组和空白对照组。结果生长抑制实验结果显示,芹菜素抑制BT474和SK-BR-3细胞生长的半数抑制浓度值分别为25.70和29.34μmol/L。EdU染色实验结果表明,不同浓度芹菜素(16、32、64μmol/L)单药作用于SK-BR-3和BT474细胞后,与对照组比较差异均有统计学意义,F值分别为99.25和309.10,均P<0.001。芹菜素(16、32、64μmol/L)与曲妥珠单抗(16.67nmol/L)联合处理SK-BR-3和BT474细胞后,与芹菜素组EdU阳性率比较,差异均有统计学意义,F值分别为145.60和110.10,均P<0.001;芹菜素+曲妥珠单抗联合用药组与曲妥珠单抗组EdU阳性率比较差异有统计学意义,F值分别为48.38和66.46,均P<0.001。蛋白质印迹结果显示,芹菜素组、曲妥珠单抗组和芹菜素+曲妥珠单抗联合用药组p-EGFR蛋白相对表达量分别为(81.75±1.08)%、(45.84±1.71)%和(17.06±1.06)%,F=2093.00,P<0.001;p-HER2蛋白相对表达量分别为(55.09±6.23)%、(41.33±2.48)%和(8.87±0.20)%,F=604.60,P<0.001;p-HER3蛋白相对表达量分别为(67.78±1.94)%、(17.89±0.84)%和(12.06±0.81)%,F=2749.00,P<0.001;p-Akt蛋白相对表达量分别为(46.85±13.96)%、(42.35±2.71)%和(23.41±0.24)%,F=85.73,P<0.001。结论芹菜素与曲妥珠单抗可协同抑制HER2Objective To explore the anti-tumor effect and mechanism of apigenin combined with trastuzumab on human epidermal growth factor receptor 2(HER2)positive breast cancer cell lines BT474 and SK-BR-3.Methods The binding activity of trastuzumab to human breast cancer cells was detected by flow cytometry;Cell counting kit(CCK)-8 was used to detect the growth inhibitory activity of apigenin,trastuzumab monotherapy,and their combination on BT474 and SK-BR-3cells;EdU staining method was used to detect the effects of apigenin,trastuzumab monotherapy,and their combination on DNA replication in BT474 and SK-BR-3 cells;Protein blotting was used to detect the effects of apigenin,trastuzumab monotherapy,and their combination on the expression levels of epidermal growth factor receptor(EGFR),HER2,HER3,Akt,and their respective phosphorylated proteins in BT474 and SK-BR-3 cells.The experiment was divided into apigenin group,trastuzumab group,apigenin+trastuzumab combination group,control group,and blank control group.Results The results of the growth inhibition experiment showed that the IC_(50)values of apigenin inhibiting the growth of BT474 and SK-BR-3 cells were 25.70 and 29.34μmol/L,respectively.The EdU staining experiment results showed that after different concentrations of apigenin(16,32,64μmol/L)were applied to SK-BR-3 and BT474 cells,and there were statistically significant differences compared to the control group,with F values of 99.25 and 309.10,both P<0.001.After treating SK-BR-3 and BT474 cells with a combination of apigenin and trastuzumab,there was a statistically significant difference in the positive rate of EdU compared to the apigenin group,with F values of 145.60 and 110.10,respectively,all P<0.001;There was a statistically significant difference in the positive rate of EdU compared to the trastuzumab group,with F values of 48.38 and 66.46,respectively,both P<0.001.The protein blot results showed that the relative expression levels of p-EGFR protein in the apigenin group,trastuzumab group,and apigenin+trast

关 键 词：乳腺癌 曲妥珠单抗 芹菜素 人表皮生长因子受体2 

分 类 号：R737.9[医药卫生—肿瘤]