[目的]探究不同浓度的黑蒜提取物(black garlic extract,BGE)对人乳腺癌细胞系-7(human breast cancer cell line-7,MCF-7)生长的影响,并进一步探讨其作用的机制.[方法]通过应用噻唑蓝法(methyl thiazolyl tetrazolium,MTT)、5-溴脱氧尿嘧啶核苷(bromo-deoxyuridine,BrDU)法检测不同浓度(25、50、100 mg/mL)BGE对于乳腺癌细胞增殖的影响.应用流式细胞术(Annexin V-FITC/PI双染色法)检测不同浓度BGE对于乳腺癌细胞凋亡的影响.应用蛋白质印迹法(Western blot)检测磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)、p-PI3K 蛋白激酶 B(Akt)、p-Akt、哺乳动物雷帕霉素靶蛋白(mam-malian target of rapamycin,mTOR)、p-mTOR蛋白的表达.应用SPSS 26软件进行单因素方差分析.[结果]MTT结果表明,经不同浓度BGE作用后,24 h BGE 25、50、100 mg/mL实验组细胞增殖率(64.14％±1.52％,56.06％±3.12％,53.39％±1.62％)低于对照组(71.79％±2.73％)(P＜0.01),48 h 实验组细胞增殖率(81.23％±3.45％,74.71％±3.58％,69.42％±2.06％)低于对照组(89.28％±1.44％)(P＜0.01),72 h实验组细胞增殖率(87.38％±3.65％,82.96％±8.46％,76.69％±6.60％)低于对照组(95.12％±3.98％)(P＜0.01).流式细胞术检测发现BGE 25、50、100 mg/mL实验组细胞凋亡率(9.57％±0.97％,15.23％±1.21％,17.42％±1.89％)高于对照组(5.53％±0.52％)(P＜0.01).Western blot 法检测发现实验组磷酸化的PI3K、Akt与mTOR蛋白的表达与对照组相比均受到抑制(P均＜0.01).[结论]BGE对乳腺癌细胞增殖有抑制作用,对其凋亡有促进作用,其作用机制可能与PI3K/Akt/mTOR通路相关.
Black Garlic Extract Promotes Apoptosis in Breast Cancer Cells Through PI3K/Akt/mTOR Pathway
[Objective]To investigate the effects and mechanism of black garlic extract(BGE)on the growth of human breast cancer cells.[Methods]Human breast cancer MCF-7 cells were treated with BGE at con-centration of 25,50 and 100 mg/mL,respectively.The proliferation and apoptosis of MCF-7 cells were de-tected by MTT,BrDU and flow cytometry(Annexin V-FITC/PI double staining)methods,respectively.The expression of phosphoinositide 3-kinase(PI3K),p-PI3K,protein kinase B(Akt),p-Akt,mammalian target of rapamycin(mTOR),and p-mTOR was detected by Western blot.One-way analysis of variance(ANOVA test)was applied with SPSS 26 software.[Results]MTT assay showed that the proliferation rate MCF-7 cells treated with BGE(25,50 and 100 mg/mL)was lower than that of the control group(for 24 h:64.14％±1.52％,56.06％±3.12％and 53.39％±1.62％vs 71.79％±2.73％,P＜0.01;for 48 h:81.23％±3.45％,74.71％±3.58％and 69.42％±2.06％vs 89.28％±1.44％,P＜0.01;for 72 h:87.38％±3.65％,82.96％±8.46％and 76.69％±6.60％vs 95.12％±3.98％,P＜0.01).Flow cytometry showed that the apoptosis rate of the BGE groups was higher than that of the control group(for 48 h:9.57％±0.97％,15.23％±1.21％and 17.42％±1.89％vs 5.53％±0.52％,P＜0.01).Western blot showed that expression of phosphorylated PI3K,Akt and mTOR pro-teins in the BGE groups were all inhibited compared with the control group(all P＜0.01).[Conclusion]BGE may inhibit proliferation and promote apoptosis of breast cancer cell through the PI3K/Akt/mTOR pathway.
black garlic extractbreast cancer cellsproliferationapoptosis
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