瑞香狼毒提取物抗乳腺癌MCF-7细胞多药耐药的研究  

Stellera chamaejasme extract against multidrug resistance of breast cancer cell line MCF-7

作　　者：崔羲和 曾蕊 臧远龙 杨庆[1] 朱晓新[1] 王娅杰[1] CUI Xi-he;ZENG Rui;ZANG Yuan-long;YANG Qing;ZHU Xiao-xin;WANG Ya-jie(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国中药杂志》2023年第9期2360-2367,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174030);中国中医科学院科技创新工程项目(CI2021B015);中央级公益性科研院所基本科研业务费项目(ZXKT20026);国家“重大新药创制”科技重大专项(2017ZX09301012002)。

摘　　要：该研究将探索瑞香狼毒提取物(SCE)抗乳腺癌多药耐药的作用及相关机制。以乳腺癌化疗敏感细胞株MCF-7及阿霉素耐药细胞株MCF-7/ADR作为实验对象,通过噻唑蓝(MTT)法检测细胞增殖活力,采用Pi单染法测定细胞周期,4′,6-二脒基-2-苯基吲哚(DAPI)染色、流式细胞术检测细胞凋亡情况,单丹磺酰尸胺(MDC)染色以及GFP-LC3B-mcherry腺病毒转染检测自噬流情况,Western blot检测凋亡蛋白Bcl-2、Bax、caspase-9、caspase-3以及自噬蛋白LC3B、p62、Beclin-1的表达水平。实验结果显示,SCE能够显著抑制乳腺癌细胞敏感/耐药株的增殖,耐药因子为0.53,远低于阿霉素的耐药因子59,同时SCE处理后敏感/耐药细胞株的G0/G1期细胞比例均显著增加。另外,SCE给药后的DAPI染色表明,敏感/耐药细胞株均出现核浓缩、染色加深、核碎裂等一系列凋亡现象;流式双染结果表明SCE给药后敏感/耐药细胞株凋亡细胞比例均显著上升;Western blot结果显示SCE给药后敏感/耐药细胞株caspase-3、caspase-9、Bcl-2蛋白的表达水平均显著降低而Bax蛋白的表达水平显著升高。进一步,SCE处理后也能够使乳腺癌耐药株MDC阳性染色以及GFP-LC3B-mcherry转染后的黄色荧光斑点增多,并上调乳腺癌细胞自噬相关蛋白LC3B-Ⅱ、p62以及Beclin-1的表达水平。综上所述,SCE可能通过阻滞乳腺癌耐药细胞的细胞周期,并阻断自噬流,最终通过干预耐药细胞的凋亡抵抗而发挥抗耐药作用。This study explored the effect and underlying mechanism of Stellera chamaejasme extract(SCE)on multidrug resistance of breast cancer.The chemotherapy-sensitive breast cancer cell line MCF-7 and adriamycin(ADR)-resistant cell line MCF-7/ADR were used as experimental subjects.MTT assay was used to detect cell proliferation activity.Pi staining was used to detect the cell cycle.4′,6-Diamidino-2-phenylindole,dihydrochloride(DAPI)staining and flow cytometry were used to detect apoptosis.Dansylcadaverine(MDC)staining and GFP-LC3B-Mcherry adenovirus transfection were used to detect autophagy.The protein expression of Bcl-2,Bax,caspase-9,caspase-3,LC3B,p62,and Beclin-1 was detected by Western blot.The results showed that SCE could significantly inhibit the proliferation of both sensitive and resistant breast cancer cell lines.The drug resistance factor was 0.53,which was significantly lower than 59 of ADR.Meanwhile,the proportion of sensitive/resistant cells in the G0/G1 phase increased significantly after SCE treatment.In addition,DAPI staining showed that a series of apoptosis phenomena such as nuclear pyknosis,staining deepening,and nuclear fragmentation appeared in sensitive/resistant cell lines after SCE administration.Moreover,the results of flow cytometry double staining showed that the proportion of apoptotic cells in sensitive/resistant cell lines increased significantly after SCE administration.Besides,Western blot showed that the protein expression levels of caspase-3,caspase-9,and Bcl-2 significantly decreased and the expression level of Bax protein significantly increased in both breast cancer cell lines after SCE administration.Furthermore,SCE could also increase the positive fluorescent spots after MDC staining and yellow fluorescent spots after GFP-LC3B-mcherry transfection,and up-regulate the expression levels of autophagy-related proteins LC3B-Ⅱ,p62,and Beclin-1 in breast cancer cells.In summary,SCE may play the role of anti-multidrug resistance by blocking the cell cycle of breast cancer multidrug-

关 键 词：瑞香狼毒提取物 乳腺癌 多药耐药 自噬 凋亡 

分 类 号：R285[医药卫生—中药学]