首页|PI3k/Akt通路在大戟大枣汤抗不同分子表型乳腺癌中的调控作用

PI3k/Akt通路在大戟大枣汤抗不同分子表型乳腺癌中的调控作用

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目的 探讨 PI3k/Akt 通路在大戟大枣汤(decoction of Euphorbia fischeriana Steud。and jujuba,DEFSJ)抗雌激素受体(ER)阴性(-)和ER阳性(+)乳腺癌中的调控作用,为乳腺癌的针对性治疗提供参考。方法 制备DEFSJ提取液,采用UHPLC-Triple Quad仪进行质控分析。DEFSJ含药血清(containing serum,CS)依照体外血清药理学方法进行获取,乳腺癌(ER-)MDA-MB-453和(ER+)MCF-7细胞分别用不同浓度DEFSJ-CS干预48 h。流式细胞术检测细胞周期分布,DNA ladder实验评估细胞凋亡程度,Western blot法检测PI3k/Akt通路相关蛋白表达,实时定量聚合酶链反应(qRT-PCR)法检测FoxO3a、FoxO1a及Bim mRNA表达;激光共聚焦显微镜观察FoxO3a蛋白核转位。结果 UPLC对5批DEFSJ提取物进行了分析,结果表明制备工艺可行,质量可控,保证了药理学实验结果的准确性。体外实验中发现,DEFSJ-CS能阻断细胞在G2/M期(P<0。05,P<0。01);DNA ladder实验中呈现典型的细胞凋亡梯带;与阴性对照组比较,DEFSJ-CS能使p-PI3k、p-Akt、p-FoxO3a及p-FoxO1a蛋白表达降低(P<0。05,P<0。01),Bim 蛋白表达升高(P<0。05);使 FoxO3a、FoxO1a mRNA 表达降低(P<0。05,P<0。01),Bim mRNA 表达升高(P<0。05,P<0。01);并能使细胞中FoxO3a蛋白呈现显著的核转位变化;且各数据结果均显示DEFSJ-CS对(ER-)MDA-MB-453细胞比对(ER+)MCF-7细胞作用效果更好。结论 PI3k/Akt通路参与了大戟大枣汤抗乳腺癌作用的调控,作用效果因乳腺癌表型差异而不同。
Effects of decoction of Euphorbia fischeriana Steud.and jujuba against breast cancer of different molecular phenotypes via PI3k/Akt pathway
Objective To explore the effect of decoction of Euphorbia fischeriana Steud.and jujuba(DEFSJ)against estrogen receptor(ER)negative(-)and ER positive(+)breast cancer via the PI3k/Akt pathway,and to provide a reference for the targeted treatment of breast cancer.Methods DEFSJ extract was prepared and analyzed using UHPLC-Triple Quad.DEFSJ containing serum(CS)was prepared via a serum pharmacology method.Different concentrations of DEFSJ-CS were applied to(ER-)MDA-MB-453 and(ER+)MCF-7 breast cancer cells in vitro for 48 h.The distribution of cells in different stages of the cellular cycle was evaluated using a Flow cytometer.DNA ladder assay was used to assess the degree of apoptosis,and the expression of PI3k/Akt pathway-related proteins was evaluated by Western blot assay.The expression of FoxO3a,FoxOla,and Bim mRNA was detected by Real-time quantitative PCR.Nuclear transposition of FoxO3a protein was analysed using a confocal laser microscopy.Results Five batches of DEFSJ extract were analyzed using UPLC,and the result showed that the preparation technology was feasible and the quality was controllable,ensuring the accuracy of the pharmacological experiment result.DEFSJ-CS blocked cells in the G2/M phase(P<0.05,P<0.01).Cells treated with DEFSJ-CS displayed the typical apoptotic ladder in the DNA ladder experiment.Compared with the negative control cells,the DEFSJ-CS group cells had decreased protein expression of p-PI3k,p-Akt,p-FoxO3a,and p-FoxO1a(P<0.05,P<0.01);increased protein expression of Bim(P<0.05);decreased mRNA expression of FoxO3a and FoxO1a(P<0.05,P<0.01);increased mRNA expression of Bim(P<0.05,P<0.01);and enhanced nuclear translocation of FoxO3a protein.The data showed that DEFSJ-CS had a stronger effect on(ER-)MDA-MB-453 cells than(ER+)MCF-7 cells.Conclusions The regulatory effect of DEFSJ extract on anti-breast cancer involves the PI3k/Akt pathway,and the effect varies with phenotypic differences.

Euphorbia fischeriana Steud.MDA-MB-453 cellsMCF-7 cellsPI3k/Akt pathway

马立威、姚宏玉、陈哲、倪世宇、陈颂、李京、刘吉成

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齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔 161006

齐齐哈尔医学院药学院,黑龙江齐齐哈尔 161006

齐齐哈尔医学院公共卫生学院,黑龙江齐齐哈尔 161006

齐齐哈尔医学院附属第五医院,黑龙江大庆 163001

齐齐哈尔医学院附属第三医院,黑龙江齐齐哈尔 161099

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狼毒大戟 MDA-MB-453细胞 MCF-7细胞 PI3k/Akt通路

国家自然科学基金齐齐哈尔市科技局联合引导项目黑龙江省中医药局科技项目

82174207LHYD-202028ZHY2020-179

2024

中国比较医学杂志
中国实验动物学会,中国医学科学院医学实验动物研究所

中国比较医学杂志

CSTPCD北大核心
影响因子:0.473
ISSN:1671-7856
年,卷(期):2024.34(3)
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