圣草酚通过miR-128-3p/MAPK轴对乳腺癌细胞增殖、凋亡的影响  

Effects of Eriodictyol on Proliferation and Apoptosis of Breast Cancer Cells via miR-128-3p/MAPK Axis

作　　者：高建朝[1] 张志生[1] 张京力 李晓霞 马科[2] 冯志林 周海丰[1] 王展海[1] GAO Jianchao;ZHANG Zhisheng;ZHANG Jingli;LI Xiaoxia;MA Ke;FENG Zhilin;ZHOU Haifeng;WANG Zhanhai(Department of Breast Surgery,Zhangjiakou 075000 Hebei,China;Department of Clinical Laboratory,TThe First Affiliated Hospital of Hebei North University,Zhangjiakou 075000 Hebei,China)

机构地区：[1]河北北方学院附属第一医院乳腺外科,河北张家口075000 [2]河北北方学院附属第一医院检验科,河北张家口075000

出　　处：《中药新药与临床药理》2023年第9期1187-1194,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：张家口市重点研发计划项目(2121135D)。

摘　　要：目的圣草酚是一种重要的类黄酮,普遍存在于植物界,但少有对圣草酚的抗癌活性的报道。该研究旨在探究其对人乳腺癌(breast cancer,BC)的抗癌潜力及可能的机制。方法体外培养人乳腺癌细胞MCF-7和人乳腺上皮细胞FR2,通过MTT法检测圣草酚诱导的初始细胞毒性。再次培养MCF-7细胞,MTT法和集落形成实验评估圣草酚对MCF-7细胞增殖的影响;流式细胞术评估圣草酚对MCF-7细胞凋亡和线粒体膜电位(mitochondrial membrane potential,MMP)的作用;RT-qPCR和Western Blot法检测miR-128-3p和MAPK相关基因(p38 MAPK和HSP27)在圣草酚发挥抗癌功能中的作用。裸鼠体内异种移植模型中分析圣草酚对肿瘤生长的作用,TUNEL法检测圣草酚对细胞凋亡的影响。结果当圣草酚浓度超过12.5μmol·L^(-1)时,MCF-7细胞活力随着圣草酚浓度的增高而逐渐降低(P<0.05);圣草酚干预后,MCF-7细胞活力、集落形成数量和MMP均下调,细胞凋亡率增高(P<0.05);细胞中miR-128-3p水平增高,p38 MAPK和HSP27磷酸化水平均降低(P<0.05)。转染miR-128-3p抑制物在一定程度上可以逆转圣草酚对MCF-7细胞的影响(P<0.05);在转染miR-128-3p抑制物的基础上添加MAPK通路抑制剂可以削弱这种逆转作用(P<0.05)。此外,裸鼠体内异种移植模型实验证明了圣草酚可呈剂量依赖性降低裸鼠肿瘤组织生长,促进细胞凋亡(P<0.05)。结论圣草酚可有效抑制人乳腺癌细胞增殖并诱导细胞凋亡,这可能与其调控miR-128-3p/MAPK轴有关。Objective Eriodictyol,an important flavonoid,which is ubiquitous in the plant kingdom.However,the anticancer activity of eriodictyol has not been well reported.This study aimed to investigate its potential anticancer activity against human breast cancer(BC)and possible mechanisms.Methods Human BC cells(MCF-7)and mammary epithelial cells(FR2)were cultured in vitro,and initial cytotoxicity induced by eriodictyol was measured by MTT assay.MCF-7 cells were cultured again,and the effect of eriodictyol on the proliferation of MCF-7 cells was evaluated by MTT method and colony formation assay.The effects of eriodictyol on apoptosis and mitochondrial membrane potential(MMP)of MCF-7 cells were assessed by flow cytometry.miR-128-3p and MAPK-related genes(p38 MAPK and HSP27)were detected by RT-qPCR and Western Blot to explore their role in anticancer function of eriodictyol.The effect of eriodictyol on tumor growth was analyzed in an in vivo xenograft model,and the effect of eriodictyol on cell apoptosis was detected by TUNEL.Results When the concentration of eriodictyol exceeded12.5μmol·L^(-1),the viability of MCF-7 cells gradually decreased with the increase of eriodictyol concentration(P<0.05).After eriodictyol intervention,the MCF-7 cell viability,colony formation number and MMP level were down-regulated,but cell apoptosis rate was increased(P<0.05).The level of miR-128-3p in the cells was increased,and the phosphorylation levels of p38 MAPK and HSP27 were decreased(P<0.05).Transfection of miR-128-3p inhibitor was able to reverse the effect of eriodictyol on MCF-7 cells to a certain extent(P<0.05).The addition of MAPK pathway inhibitor and the transfection of miR-128-3p inhibitor were able to weaken the reversal effect of miR-128-3p in MCF-7 cells(P<0.05).In addition,in vivo experiments demonstrated that eriodictyol was able to dose-dependently reduce tumor tissue growth and promote cell apoptosis(P<0.05).Conclusion Eriodictyol can effectively inhibit the proliferation and induce apoptosis of BC cells,which may be rel

关 键 词：圣草酚 miR-128-3p MAPK 乳腺癌 增殖 凋亡 人乳腺癌细胞MCF-7 人乳腺上皮细胞FR2 裸鼠 

分 类 号：R285.5[医药卫生—中药学]