冬凌草甲素逆转乳腺癌MCF-7细胞对氟维司群耐药的机制研究  

Study on the mechanism of oridonin to reverse drug resistance of breast cancer cell MCF-7 to fulvestrant

作　　者：胡高波[1] 朱瑞 金湛 魏自太[1] HU Gaobo;ZHU Rui;JIN Zhan;WEI Zitai(Medical School,Quzhou College of Technology,Zhejiang Quzhou 324000,China;College of Pharmacy,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]衢州职业技术学院医学院,浙江衢州324000 [2]浙江中医药大学药学院,杭州310053

出　　处：《中国药房》2022年第17期2119-2123,共5页China Pharmacy

基　　金：衢州市科技计划项目(No.2019K41);衢州市重点创新团队资助项目(No.衢委人才[2017]6号)。

摘　　要：目的探究冬凌草甲素(Ori)逆转乳腺癌MCF-7细胞对氟维司群(Ful)耐药的机制。方法在体外诱导构建乳腺癌获得性Ful耐药细胞株MCF-7/Ful;采用MTT法检测MCF-7细胞和MCF-7/Ful细胞的相对细胞活力,Ori对MCF-7/Ful细胞的抑制率;采用CompuSyn软件分析Ori和Ful的协同效应;将MCF-7/Ful细胞随机分为空白对照组、Ful组(5μmol/L)、Ori组(8μmol/L)、Ful(5μmol/L)+Ori(8μmol/L)组,采用Western blot法检测各组细胞中磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路相关蛋白的磷酸化情况。采用MCF-7/Ful细胞制备裸鼠移植瘤耐药模型,随机分为空白对照组、Ful组(80μmol/g)、Ori组(50μmol/g)、Ful(80μmol/g)+Ori(50μmol/g)组,计算肿瘤质量和抑瘤率,验证Ori在体内的耐药逆转作用。结果MTT法检测结果显示,当Ful≥10μmol/L时,MCF-7/Ful细胞的相对细胞活力显著高于MCF-7细胞(P<0.05),耐药性显著增强;Ori对MCF-7/Ful细胞具有明显的抑制作用,与Ful联合使用对MCF-7/Ful细胞的抑制率显著升高(P<0.05或P<0.01),逆转耐药效果显著增加。Western blot法检测结果显示,与Ful组比较,Ful+Ori组细胞中PI3K、Akt磷酸化水平均显著降低(P<0.05或P<0.01)。体内实验结果显示,Ful+Ori组裸鼠的肿瘤体积和质量显著下降,抑瘤率为(63.90±4.11)%,较Ful组显著升高(P<0.01)。结论Ori可以逆转乳腺癌MCF-7细胞对Ful的耐药性,此作用可能是通过调控PI3K/Akt信号通路实现的。OBJECTIVE To explore the mechanism of oridonin(Ori)reversing the drug resistance of breast cancer cell MCF-7 to fulvestrant(Ful).METHODS Ful-resistant breast cancer cell strains MCF-7/Ful were induced and constructed in vitro.The relative cell viability of MCF-7 cells and MCF-7/Ful cells was detected by MTT assay,inhibitory rate of Ori to MCF-7/Ful cells was also detected.CompuSyn software was used to analyze the synergistic effect of Ori and Ful.MCF-7/Ful cells were randomly divided into blank control group,Ful group(5μmol/L),Ori group(8μmol/L),Ful(5μmol/L)+Ori(8μmol/L)group.The phosphorylation of phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)signaling pathway related protein in each group was detected by Western blot.MCF-7/Ful cells were used to prepare drug resistance model of transplanted tumor in nude mice,and they were randomly divided into blank control group,Ful group(80μmol/g),Ori group(50μmol/g),Ful(80μmol/g)+Ori(50μmol/g)group.The tumor weight and tumor inhibition rate were calculated,to verify the reversal effect of Ori and Ful in vivo.RESULTS MTT assay showed that when Ful≥10μmol/L,the relative cell viability of MCF-7/Ful cells was significantly higher than that of MCF-7 cells(P<0.05),and the drug resistance was significantly enhanced;Ori had a significant inhibitory effect on MCF-7/Ful cells,the inhibition rate of Ori combined with Ful to MCF-7/Ful cells was significantly increased(P<0.05 or P<0.01),and the effect of reversing drug resistance was significantly increased.The results of Western blot showed that compared with Ful group,the phosphorylation levels of PI3K and Akt protein in Ful+Ori group were significantly decreased(P<0.05 or P<0.01).In vivo results showed that the tumor volume and mass of Ful+Ori group were significantly decreased,and the tumor inhibition rate was(63.90±4.11)%,which was significantly higher than that of Ful group(P<0.01).CONCLUSIONS Ori can reverse drug resistance of breast cancer cell MCF-7 to Ful,and this effect may be through regulating PI3 K/A

关 键 词：冬凌草甲素 氟维司群 磷脂酰肌醇-3-激酶/蛋白激酶B信号通路 乳腺癌 耐药逆转 

分 类 号：R965[医药卫生—药理学] R737.9[医药卫生—药学]