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黄芪总皂苷通过LKB1/AMPK信号通路诱导乳腺癌MCF-7细胞自噬的分子机制研究
作者:张鹏1  徐洪燕2  张启文1 
单位:1. 济南市人民医院 两腺外科, 山东 济南 271100;
2. 济南市人民医院 放疗科, 山东 济南 271100
关键词:黄芪总皂苷 LKB1/AMPK信号通路 乳腺癌MCF-7细胞 自噬 
分类号:R285;R737.9
出版年·卷·期(页码):2020·48·第九期(1158-1163)
摘要:

目的:探讨黄芪总皂苷通过肝激酶B1/腺苷酸活化蛋白激酶(LKB1/AMPK)信号通路诱导乳腺癌MCF-7细胞自噬的分子机制。方法:将10 ml乳腺癌MCF-7细胞培养于含10%胎牛血清的DMEM培养液中。置于CO2培养箱为乳腺癌MCF-7细胞组;10 ml乳腺癌MCF-7细胞培养于含10%胎牛血清的DMEM培养液中,加入5 ml黄芪总皂苷(80 mg/UL)置于CO2培养箱为黄芪总皂苷低剂量组;黄芪总皂苷高剂量组加入5 ml黄芪总皂苷(160 mg·μl-1)。培养结束后,MTT法检测各组细胞活性,荧光显微镜观察自噬小体水平,倒置显微镜观察克隆形成数目,流式细胞法测定凋亡率、G1期水平,实时荧光逆转录法测定LKB1、AMPK mRNA水平,酶联免疫吸附法测定LKB1、AMPK蛋白表达水平。结果:黄芪总皂苷低、高剂量组细胞OD值、存活率水平、细胞克隆形成数目均低于乳腺癌MCF-7细胞组(P<0.05),黄芪总皂苷高剂量组OD值、存活率、细胞克隆形成数目低于黄芪总皂苷低剂量组(P<0.05)。黄芪总皂苷低、高剂量组细胞自噬小体、G1期水平、凋亡率、LKB1、AMPK mRNA、蛋白表达水平高于乳腺癌MCF-7细胞组(P<0.05),黄芪总皂苷高剂量组自噬小体、G1期水平、凋亡率、LKB1、AMPK mRNA、蛋白表达水平高于黄芪总皂苷低剂量组(P<0.05)。结论:黄芪总皂苷通过促进LKB1、AMPK mRNA及蛋白水平高表达激活LKB1/AMPK信号通路,诱导MCF-7细胞自噬。

Objective: To investigate the molecular mechanism of autophagy of breast cancer MCF-7 cells induced by total saponins of Astragalus membranaceus through liver kinase B1/AMP-activated protein kinase(LKB1/AMPK) signaling pathway. Methods: 10 mL breast cancer MCF-7 cells were cultured in DMEM medium containing 10% fetal bovine serum in CO2incubator as breast cancer MCF-7 cells group, 10 mL breast cancer MCF-7 cells were cultured in DMEM medium containing 10% fetal bovine serum plus 5 mL Astragaloside (80 mg/UL) in CO2 incubator as low dose of total saponins of Astragaloside group, and 5 mL astragaloside (160 mg/UL) was added in high dose of total saponins of Astragaloside group. After culture, MTT assay was used to detect the cell viability, fluorescence microscopy was used to determine the level of autophagosomes, inverted microscopy was used to observe the number of clones, flow cytometry was used to determine the apoptotic rate and G1 phase, real-time fluorescence reverse transcription was used to determine the levels of LKB1 and AMPK mRNA, and enzyme-linked immunosorbent assay was employed to detect the expression levels of LKB1 and AMPK protein.Results: The OD value, survival rate and number of cell clones in low and high dose groups were lower than those in breast cancer MCF-7 cell group (P<0.05). The OD value, survival rate and number of cell clones in high dose group were lower than those in low dose group (P<0.05). The levels of autophagosome, G1 phase, apoptotic rate, LKB1, AMPK mRNA and protein expression in low and high dose groups were higher than those in breast cancer MCF-7 cell group (P<0.05). The levels of autophagosome, G1 phase, apoptotic rate, LKB1, AMPK mRNA and protein expression in high dose group were higher than those in low dose group (P<0.05).Conclusion: The total saponins of Astragalus induce autophagy of MCF-7 cells by promoting high expression of LKB1, AMPK mRNA and protein to activate LKB1/AMPK signaling pathway.

参考文献:

[1] 吴好好,林栩.黄芪总苷对足细胞损伤保护作用的研究进展[J].右江医学,2016,44(6):707-709.
[2] 杜杰.黄芪总苷和人参皂苷降低缺血性脑卒中患者脑脊液中谷氨酸、磷酸化[J].东南大学学报(医学版),2017,36(5):828-832.
[3] 丁海霞,樊成虎,王玉泉,等.黄芪总苷对脊髓型颈椎病大鼠模型椎间盘细胞因子IL-6、TNF-α的影响[J].西部中医药,2017,30(7):20-25.
[4] 宋元春,赛依帕,高霞,等.黄芪总苷对阿霉素诱导损伤足细胞的保护作用[J].西部中医药,2018,31(3):19-22.
[5] 刘亚静,刘长江,宁佳,等.黄芪总苷氯化钠注射液治疗冠心病的临床疗效观察[J].河北医药,2017,39(19):2989-2991.
[6] LI Y C,QIAO J Y,WANG B Y,et al.Paeoniflorin ameliorates fructose-induced insulin resistance and hepatic steatosis by activating LKB1/AMPK and AKT pathways[J].Nutrients,2018,10(8):E1024.
[7] LI N S,ZOU J R,LIN H,et al.LKB1/AMPK inhibits TGF-β1 production and the TGF-β signaling pathway in breast cancer cells[J].Tumor Biol,2016,37(6):8249-8258.
[8] SUN A,LI C,CHEN R,et al.GSK-3β controls autophagy by modulating LKB1-AMPK pathway in prostate cancer cells[J].Prostate,2016,76(2):172-183.
[9] BAKULA D,MVLLER A J,ZULEGER T,et al.WIPI3 and WIPI4β-propellers are scaffolds for LKB1-AMPK-TSC signalling circuits in the control of autophagy[J].Nat Commun,2017,8(16):15637-15642.
[10] KAN J Y,YEN M C,WANG J Y,et al.Nesfatin-1/Nucleobindin-2 enhances cell migration,invasion,and epithelial-mesenchymal transition via LKB1/AMPK/TORC1/ZEB1 pathways in colon cancer[J].Oncotarget,2016,7(21):31336-31349.
[11] ZHOU Y,TONG X,REN S,et al.Synergistic anti-liver fibrosis actions of total astragalus saponins and glycyrrhizic acid via TGF-β1/Smads signaling pathway modulation[J].J Ethnopharmacol,2016,190(15):83-90.
[12] LIU Q,ZHANG L,SHAN Q,et al.Total flavonoids fromAstragalusalleviate endothelial dysfunction by activating the Akt/eNOS pathway[J].J Int Med Res,2018,46(6):2096-2103.
[13] XU E,TIAN S,MIAO M,et al.Effect of different component ratio of Astragalus total saponins and Verbena total glycosides on the cerebral infarction area and serum biochemical indicators in the focal cerebral ischemia-reperfusion rat model[J].Saudi Pharm J,2017,25(4):660-665.
[14] ZHANG X P,LI Y D,LUO L L,et al.Astragalus saponins and liposome constitute an efficacious adjuvant formulation for cancer Vaccines[J].Cancer Biother Radiopharm,2018,33(2):25-31.
[15] MAURO L,NAIMO G D,GELSOMINO L,et al.Uncoupling effects of estrogen receptor αon LKB1/AMPK interaction upon adiponectin exposure in breast cancer[J].Faseb Journal,2018,32(8):4343-4355.
[16] SHOHREH M,POWER J H T,CHATAWAY T K,et al.A comparison of LKB1/AMPK/mTOR metabolic axis response to global ischaemia in brain,heart,liver and kidney in a rat model of cardiac arrest[J].BMC Cell Biol,2018,19(1):7.
[17] ZHENG X,CHI J,ZHI J,et al.Aurora-A-mediated phosphorylation of LKB1 compromises LKB1/AMPK signaling axis to facilitate NSCLC growth and migration[J].Oncogene,2017,37(4):874-889.
[18] LEE J,YANG G,KIM Y J,et al.Hydrogen-rich medium protects mouse embryonic fibroblasts from oxidative stress by activating LKB1-AMPK-FoxO1 signal pathway[J].Biochem Biophys Res Commun,2017,491(3):733-739.

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