PARP抑制剂、盐霉素、依维莫司、舒尼替尼对乳腺癌BT-20干细胞抑制作用的初步研究

发布时间：2018-04-18 16:46

  本文选题：PARP抑制剂 + 乳腺癌　； 参考：《新乡医学院》2015年硕士论文

【摘要】：背景肿瘤干细胞学说认为,恶性肿瘤中的干细胞亚群具有自我更新和多项分化的能力,是恶性肿瘤复发、转移的根本原因。聚腺苷酸二磷酸核糖转移酶(poly ADP-ribose polymerase,PARP)在细胞中的主要功能为对损伤、断裂的DNA链进行识别和修复,临床上对乳腺癌进行的放疗、化疗的主要作用机制即为损伤和破坏肿瘤细胞的DNA,从而造成肿瘤细胞的死亡,PARP被认为是导致恶性肿瘤对放疗、化疗产生抗性的重要原因,所以它也成为了治疗乳腺癌的一个新靶点。盐霉素是一种已经经实验证实的对乳腺癌干细胞具有杀伤作用的药物。依维莫司是一种哺乳动物雷帕霉素靶蛋白(mTOR)抑制剂,临床主要用于器官移植后的免疫抑制及抗肿瘤应用。舒尼替尼是一种多靶点酪氨酸激酶抑制剂,临床上主要用于晚期肾细胞癌(renal cell carcinoma ,RCC)和胃肠道间质瘤(gastrointestinal stromal tumor, GIST)的治疗。目的分别对经过PARP抑制剂、盐霉素、依维莫司、舒尼替尼处理后的乳腺癌细胞和未经过药物处理的空白组细胞中的干细胞亚群进行标记,对比两组细胞中干细胞群所占比例,探究各种药物对腺癌干细胞有无抑制作用。方法取三阴性乳腺癌细胞株BT-20细胞,用不同浓度的5种药物分别对BT-20细胞进行处理,计算出各种药物的半数抑制浓度,从中挑选出各自药物的合适浓度(对BT-20细胞的抑制率为10%左右),用5种药物对BT-20细胞再次进行处理,ALDEFLUOR试剂标记出加药细胞组和未加药细胞的ALDH1+细胞,利用流式细胞仪检测并对比两组细胞间ALDH1阳性细胞比例,检验药物对干细胞亚群的抑制作用。结果Veliparib.Olaparib.Iniparib.盐霉素、依维莫司、舒尼替尼的ICso分别为32.23、38.65、35.12、7.21、5.73、7.85μmol/L。未加药组细胞中的ALDH1+细胞比例为(19.34±1.15)%,实验组细胞中的ALDHl+细胞比例为(盐霉素,1μmol/L)(10.87±2.24)%；(盐霉素,2μmol/L)(6.33±1.52)%；(Olaparib,10μmol/L)(8.45±0.56)%； (Veliparib,10μmol/L)(13.27±2.14)%；(Iniparib,10μmol/L)(6.83±2.67)%；(依维莫司,1μmol/L)(9.82±1.74)%；(舒尼替尼,1μmol/L)(7.87±2.52)%.实验组细胞的ALDH1+细胞比例显著降低,(P0.05)。结论经PARP抑制剂(Veliparib、Olaparib、Iniparib)、盐霉素、依维莫司、舒尼替尼作用后的BT-20细胞中ALDH1+细胞比例明显降低,以上药物对乳腺癌细胞株BT-20中的干细胞亚群有直接抑制作用。Primary research of the inhibition of PARP inhibitor,Salinomycin, Everolimus and Sunitinib on breast cancer BT-20 stem cells
[Abstract]:Background: according to the theory of tumor stem cells, stem cell subsets in malignant tumors have the ability of self-renewal and multi-differentiation, which is the fundamental cause of recurrence and metastasis of malignant tumors.The main function of polyadenylate diphosphate ribonucleosyltransferase (ADP-ribose) in cells is to recognize and repair the damaged and broken DNA chains, and to treat breast cancer with radiotherapy.The main mechanism of chemotherapy is to damage and destroy the DNA of tumor cells, resulting in the death of tumor cells. PARP is considered to be an important cause of cancer resistance to radiotherapy and chemotherapy.So it has become a new target for breast cancer.Salinomycin is a proven drug that can kill breast cancer stem cells.Evimostr is a mammalian rapamycin target protein mTOR-inhibitor, which is mainly used for immunosuppressive and antitumor applications after organ transplantation.Sulnitinib is a multitarget tyrosine kinase inhibitor, which is mainly used in the treatment of advanced renal cell carcinoma (RCC) and gastrointestinal stromal tumor (GIST).Objective to label the stem cell subsets in breast cancer cells treated with PARP inhibitor, salinomycin, ivimostatin and sunitinib, and to compare the proportion of stem cell groups in the two groups.To explore the inhibitory effect of various drugs on adenocarcinoma stem cells.Methods three negative breast cancer cell line BT-20 cells were treated with different concentrations of 5 drugs to calculate the half inhibitory concentration of each drug.The appropriate concentration of each drug was selected (the inhibition rate on BT-20 cells was about 10%), and ALDH1 cells were labeled with ALDEFLUOR reagent in addition cells and untreated cells with 5 kinds of drugs, and then the cells were labeled with ALDEFLUOR reagent, and the cells were labeled with ALDEFLUOR reagent, and the ALDH1 cells were labeled with ALDEFLUOR reagent.Flow cytometry was used to detect and compare the ratio of ALDH1 positive cells between the two groups, and the inhibitory effect of drugs on stem cell subsets was tested.Results Veliparib.Olaparib.Iniparib.The ICso of salinomycin, ivimostr and sulnitinib were 32.2338.65 渭 mol / L, 35.127.21 and 5.73 渭 mol / L.鏈�鍔犺嵂缁勭粏鑳炰腑鐨凙LDH1 缁嗚優姣斾緥涓，

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