大豆苷元增强多西紫杉醇体外杀伤三阴性乳腺癌作用研究
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摘要
目的:研究大豆苷元通过调节BRCA1表达逆转多西紫杉醇耐药的具体机制。方法:应用三阴性人乳腺癌细胞系MDA-MB-231、SUM-1315作为研究对象,通过MTT法评估分析不同治疗组的抑瘤率,应用流式细胞仪检测各治疗组细胞的凋亡情况;应用western-blot分析各治疗组BRCA1及凋亡相关蛋白的表达情况。结果:研究发现大豆苷元、多西紫杉醇及联合组对乳腺癌MDA-MB-231、SUM1315细胞的抑制呈浓度依赖的原则。并且BRCA1表达的细胞系MDA-MB-231比BRCA1突变的细胞系SUM1315抑瘤率更高,两个细胞系不同治疗组分别与对照组相比较,凋亡率明显升高,均具有统计学意义;且随着大豆苷元的加入,可使多西紫杉醇的凋亡率显著增加,并且可减少多西紫杉醇的用量。在SUM-1315细胞系,大豆苷元逆转多西紫杉醇耐药,增加凋亡比率更为显著。大豆苷元可使BRCA1突变型的三阴性乳腺癌细胞系SUM-1315表达BRCA1,并具有浓度依赖性。对各组细胞的凋亡蛋白的变化进行免疫印迹分析表明大豆苷元联合多西紫杉醇可通过调节caspase依赖的凋亡通路活性来增强多西紫杉醇的凋亡作用,从而增强乳腺癌细胞杀伤。结论:大豆苷元可通过调节BRCA1表达量,协同增强多西紫杉类药物杀伤三阴性乳腺癌细胞,促进凋亡进而逆转化疗耐药。
Objective: To study the specific mechanism of daidzein to reverse docetaxel resistance by regulating the expression of BRCA1. Methods: The tri-negative human breast cancer cell line MDA-MB-231 and SUM-1315 were used as the study, and the MTT method was used to evaluate the rate of tumor grow inhibition during different treatment groups, and the cell apoptosis was detected by flow cytometry. Apoptosis related proteins expressions were analyzed by western-blot. Results: We found that the inhibition rate of the daidzein, docetaxel and combined groups were concentration-dependent in the MDA-MB-231 and SUM1315 cells. And the inhibitory rate in MDA-MB-231 is higher than SUM1315. Different treatment groups compared with control group in both two cell lines, apoptosis rate increased significantly. And all groups have statistically significant. With the addition of daidzein, the apoptosis rate of docetaxel groups can be significantly increased, at the same time, the dosage of docetaxel can be reduced. In the SUM-1315 cell line, daidzein can reverse docetaxel resistance and increase apoptosis ratios. Daidzein can change the expression of BRCA1 accroding to the concentration dependence in the tri-negative breast cancer cell line. The change of apoptosis protein immunoblot analysis in each cell line showed that daidzein and docetaxel by adjusting apoptotic pathways of caspase dependent activity to enhance the apoptosis effect of docetaxel, thus improve cytotoxicity. Conclusions: This paper illustrates the daidzein enhance docetaxel killing activity and reverse the drug resistance mechanism by regulating the expression of BRCA1 in tri-negative breast cancer cells.
Objective: To study the specific mechanism of daidzein to reverse docetaxel resistance by regulating the expression of BRCA1. Methods: The tri-negative human breast cancer cell line MDA-MB-231 and SUM-1315 were used as the study, and the MTT method was used to evaluate the rate of tumor grow inhibition during different treatment groups, and the cell apoptosis was detected by flow cytometry. Apoptosis related proteins expressions were analyzed by western-blot. Results: We found that the inhibition rate of the daidzein, docetaxel and combined groups were concentration-dependent in the MDA-MB-231 and SUM1315 cells. And the inhibitory rate in MDA-MB-231 is higher than SUM1315. Different treatment groups compared with control group in both two cell lines, apoptosis rate increased significantly. And all groups have statistically significant. With the addition of daidzein, the apoptosis rate of docetaxel groups can be significantly increased, at the same time, the dosage of docetaxel can be reduced. In the SUM-1315 cell line, daidzein can reverse docetaxel resistance and increase apoptosis ratios. Daidzein can change the expression of BRCA1 accroding to the concentration dependence in the tri-negative breast cancer cell line. The change of apoptosis protein immunoblot analysis in each cell line showed that daidzein and docetaxel by adjusting apoptotic pathways of caspase dependent activity to enhance the apoptosis effect of docetaxel, thus improve cytotoxicity. Conclusions: This paper illustrates the daidzein enhance docetaxel killing activity and reverse the drug resistance mechanism by regulating the expression of BRCA1 in tri-negative breast cancer cells.
引文
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[22]Van der Woude H,Ter Veld MG,Jacobs N,et al.The stimulation of cell proliferation by quercetin is mediated by the estrogen receptor[J].Mol Nutr Food Res,2005,49(8):763-771
[2]Leclère B,MoliniéF,Trétarre B,et al.Trends in incidence of breast cancer among women under 40 in seven European countries:AGRELL cooperative study[J].Cancer Epidemiol,2013,37:544-549
[3]Yeh J,Chun J,Schwartz S,et al.Clinical Characteristics in Patients with Triple Negative Breast Cancer[J].Int J Breast Cancer,2017:1796145
[4]Ademuyiwa FO,Gao F,Hao L,et al.US breast cancer mortality trends in young women according to race[J].Cancer,2015,121:1469-1476
[5]Lafarge S,Sylvain V,Ferrara M,et al.Inhibition of BRCA1 leads to increased chemoresistance to microtubule-interfering agents,an effect that involves the JNK pathway[J].Oncogene,2001,20(45):6597-6606
[6]Quinn JE,Kennedy RD,Mullan PB,et al.BRCA1 functions as a differential modulator of chemotherapy-induced apoptosis[J].Cancer Res,2003,63(19):6221-6228
[7]Bernard-Gallon DJ,Maurizis JC,Rio PG,et al.Influence of genistein and daidzein on Brca1 protein levels in human breast cell lines[J].JNatl Cancer Inst,1998,90(11):862-863
[8]Anna Cabanes,Mingyue Wang,Susan Olivo,et al.Prepubertal estradiol and genistein exposures up-regulate BRCA1 mRNA and reduce mammary tumorigenesis[J].Carcinogenesis,2004,25(5):741-748
[9]P Tassone,P Tagliaferri,A Perricelli,et al.BRCA1 expression modulates chemosensitivity of BRCA1-defective HCC1937 human breast cancer cells[J].British Journal of Cancer,2003,88:1285-1291
[10]Sikov WM,Berry DA,Perou CM,et al.Impact of the addition of carboplatin and/or bevacizumab to neoadjuvant once-per-week paclitaxel followed by dose-dense doxorubicin and cyclophosphamide on pathologic complete response rates in stage II to III triple-negative breast cancer:CALGB 40603(Alliance)[J].J Clin Oncol,2015,33:13-21
[11]Lee A,Djamgoz MBA.Triple negative breast cancer:Emerging therapeutic modalities and novel combination therapies[J].Cancer Treat Rev,2017,(62):110-122
[12]Telli ML,Jensen KC,Vinayak S,et al.Phase II Study of Gemcitabine,Carboplatin,and Iniparib As Neoadjuvant Therapy for Triple-Negative and BRCA1/2 Mutation-Associated Breast Cancer With Assessment of a Tumor-Based Measure of Genomic Instability:Pr ECOG0105[J].J Clin Oncol,2015,33:1895-1901
[13]Petrelli F,Coinu A,Borgonovo K,et al.The value of platinum agents as neoadjuvant chemotherapy in triple-negative breast cancers:a systematic review and meta-analysis[J].Breast Cancer Res Treat,2014,144:223-232
[14]Pfeffer CM,Ho BN,Singh ATK.The Evolution,Functions and Applications of the Breast Cancer Genes BRCA1 and BRCA2[J].Cancer Genomics Proteomics,2017,14(5):293-298
[15]Stoppa-Lyonnet D.The biological effects and clinical implications of BRCA mutations:where do we go from here?[J].Eur J Hum Genet,2016,24(Suppl 1):S3-9
[16]Kais Z,Chiba N,Ishioka C,et al.Functional differences among BR-CA1 missense mutations in the control of centrosome duplication[J].Oncogene,2012,31(6):799-804
[17]Mazar J,Rosado A,Shelley J,et al.The long non-coding RNA GAS5differentially regulates cell cycle arrest and apoptosis through activation of BRCA1 and p53 in human neuroblastoma[J].Oncotarget,2017,8(4):6589-6607
[18]Masuda T,Xu X,Dimitriadis EK,et al."DNA Binding Region"of BRCA1 Affects Genetic Stability through modulating the Intra-S-Phase Checkpoint[J].Int J Biol Sci,2016,12(2):133-143
[19]Alipour S,Jafari-Adli S,Eskandari A.Benefits and harms of phytoestrogen consumption in breast cancer survivors[J].Asian Pac J Cancer Prev,2015,16(8):3091-3396
[20]Li L,Chen X,Liu CC,et al.Phytoestrogen Bakuchiol Exhibits In Vitro and In Vivo Anti-breast Cancer Effects by Inducing S Phase Arrest and Apoptosis[J].Front Pharmacol,2016,7:128
[21]De Giorgi U,Rosti G,Frassineti L,et al.High-dose chemotherapy for triple negative breast cancer[J].Ann Oncol,2007,18(1):202
[22]Van der Woude H,Ter Veld MG,Jacobs N,et al.The stimulation of cell proliferation by quercetin is mediated by the estrogen receptor[J].Mol Nutr Food Res,2005,49(8):763-771
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