青蒿素对线粒体损伤诱导的乳腺癌细胞凋亡的影响  

Effects of artemisinin on mitochondrial damage-induced apoptosis of breast cancer cells

作　　者：郭海燕 王红钰[1] 李晓英[1] 孙琪 李玉红[1] GUO Hai-yan;WANG Hong-yu;LI Xiao-ying;SUN Qi;LI Yu-hong(Department of Oncology,Tangshan Workers y Hospital,Tangshan 063000,Hebei Province,China)

机构地区：[1]唐山市工人医院肿瘤外科,河北唐山063000

出　　处：《中国临床药理学杂志》2022年第6期508-512,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的 研究青蒿素(ART)通过线粒体损伤途径诱导乳腺癌细胞凋亡的作用机制。方法 不同浓度的青蒿素(0,20,40,60μmol·L^(-1))处理乳腺癌MCF-7细胞,用噻唑蓝法检测细胞存活率;并将MCF-7细胞分为4组:Control组(不做处理)、ART组(60μmol·L^(-1)青蒿素处理)、ART+anti-miR-NC(60μmol·L^(-1)青蒿素处理+转染anti-miR-NC)、ART+anti-miR-483-5p(60μmol·L^(-1)青蒿素处理+转染anti-miR-483-5p)。用流式细胞术检测各组细胞凋亡率;用蛋白质印迹法检测各组细胞Cl-caspase-3、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞色素C(Cyto-C)和细胞凋亡诱导因子(AIF)蛋白表达情况;用实时荧光定量逆转录聚合酶链反应(qRT-PCR)发检测各组细胞miR-483-5p表达情况。结果 Control组、ART组、ART+anti-miR-NC组和ART+anti-miR-483-5p组的细胞凋亡率分别为(6.01±0.87)%,(19.06±1.67)%,(20.11±2.04)%,(10.37±1.07)%;线粒体Cyto-C蛋白表达量分别为0.91±0.09,0.39±0.05,0.42±0.06,0.85±0.07;线粒体AIF蛋白表达量分别为0.88±0.10,0.38±0.05,0.35±0.06,0.78±0.09;细胞质Cyto-C蛋白表达量分别为0.41±0.05,0.73±0.07,0.81±0.09,0.46±0.06;细胞质AIF蛋白表达量分别为0.31±0.03,0.90±0.09,0.86±0.10,0.42±0.06;各组miR-483-5p表达量分别为1.00±0.06,2.39±0.29,2.46±0.20,1.23±0.08。以上指标:Control组与ART组比较,差异均有统计学意义(均P<0.05),ART+anti-miR-NC组与ART+anti-miR-483-5p组比较,差异均有统计学意义(均P<0.05)。结论 青蒿素可通过miR-483-5p激活线粒体损伤进而诱导MCF-7细胞凋亡。Objective To study the mechanism of artemisinin (ART)on apoptosis of breast cancer cells through mitochondrial injury pathway.Methods Breast cancer MCF-7 cells were treated with different concentrations of artemisinin (0,20,40,60μmol·L^(-1)),MTT assay was used to detect the cell survival rate,MCF-7 cells were divided into four groups:control group (untreated),ART group (60μmol·L^(-1)artemisinin treatment),ART+anti-miR-NC (60μmol·L^(-1)artemisinin treatment+transfection anti-miR-NC),ART+antimiR-483-5 p (60μmol·L^(-1)artemisinin treatment+transfection anti-miR-483-58 p).The apoptosis rate of each group was detected by flow cytometry;the expression of Cl-caspase-3,B-cell lymphoma-2 (Bcl-2),Bcl-2-associated X protein (Bax),cytochrome (Cyto-C) and apoptosisinducing factor (AIF) were detected by Western blot;the expression of miR-483-5p was detected by quantitative real-time polymerase chain reaction (qRT-PCR).Results The apoptosis rates of the control group,ART group,ART+anti-miR-NC group and ART+anti-miR-483-5p group were (6.01±0.87)%,(19.06±1.67)%,(20.11±2.04)%and (10.37±1.07)%,the expression levels of mitochondrial Cyto-C protein were 0.91±0.09,0.39±0.05,0.42±0.06 and 0.85±0.07,mitochondrial AIF protein expression were 0.88±0.10,0.38±0.05,0.35±0.06,0.78±0.09,Cyto-C protein expression levels in cytoplasm were 0.41±0.05,0.73±0.07,0.81±0.09 and 0.46±0.06,the expression of AIF protein in cytoplasm were 0.31±0.03,0.90±0.09,0.86±0.10,and 0.42±0.06,the expression levels of miR-483-5p were 1.00±0.06,2.39±0.29,2.46±0.20and 1.23±0.08.The differences of above indicators between the control group and the ART group were statistically significant (all P<0.05),and the differences between the ART+anti-miR-NC group and the ART+anti-miR-483-5p group were statistically significant (all P<0.05).Conclusion Artemisinin can induce apoptosis of MCF-7 cells by activating mitochondrial damage through miR-483-5p.

关 键 词：青蒿素 线粒体损伤 微小RNA-483-5p 乳腺癌 

分 类 号：R28[医药卫生—中药学]