牛血清白蛋白胶囊释放水凝胶诱导癌细胞死亡的研究

发布时间：2018-06-25 22:14

本文选题：癌症 + 凝胶治疗　；参考：《哈尔滨工业大学》2017年硕士论文

【摘要】：癌症一直是危害人类生命健康的一大杀手。如何安全有效的治疗癌症一直是科学家们研究的重点。临床医学上最常用的方法是化疗和放疗,但是都伴随着严重的副作用。近年来,发现了一些新的治疗癌症的方法,例如基因治疗,光热治疗等。本论文中,我们将提出一种新的治疗癌症的方法“凝胶治疗方法”,该方法在不利用抗癌药物的前提下,可以选择性地利用肿瘤细胞内部高浓度的谷胱甘肽(GSH),在不需要外界刺激的条件下诱导癌细胞的死亡,避免了对正常组织细胞的副作用。本文主要是利用牛血清白蛋白胶囊将海藻酸钙凝胶(CaAlg)运输进入癌细胞,在癌细胞内高浓度GSH作用下释放凝胶,引起细胞质的原位凝胶化,从而杀死癌细胞。具体工作如下:(1)牛血清白蛋白胶囊的构筑:利用聚N-异丙基丙烯酰胺(PNIPAAm)和氨基化的牛血清白蛋白(BSA-NH2)反应得到的蛋白质-聚合物耦合体(BSA-NH2/PNIPAAm)作为基元,通过皮克林微乳液法制备蛋白质胶囊。通过调节基元浓度、超声时间和超声功率等条件制得了结构稳定,尺寸可控的(1.32μm左右)蛋白质胶囊。(2)装载海藻酸钠(NaAlg)的蛋白质胶囊的制备与表征:在蛋白质胶囊中装载海藻酸钠(NaAlg),通过调控钙离子浓度的方法使胶囊内部生成海藻酸钙凝胶,通过SEM、TEM证明了凝胶的生成;然后利用谷胱甘肽诱导胶囊释放凝胶,通过显微镜观察到蛋白质胶囊装载海藻酸钠,生成凝胶和释放凝胶的过程。(3)利用AFM技术原位检测细胞内部凝胶化过程:利用AFM测试并计算细胞的杨氏模量,通过细胞杨氏模量的变化反应了细胞的生命状态,进一步证明了凝胶形成和凝胶释放的过程。(4)体外实验:首先研究培养时间和胶囊浓度对细胞内吞作用的影响,然后对人类肝癌细胞(HepG2)和小鼠胚胎成纤维组织细胞(NIT 3T3)分别进行细胞毒性试验(MTT),通过MTT实验检了测细胞活性,对比发现凝胶法使得癌细胞的活性明显下降,而对正常细胞没有太大的影响,说明该方法可以选择性地杀死癌细胞。
[Abstract]:Cancer has been a major killer of human life and health. How to treat cancer safely and effectively has been the focus of scientists. The most commonly used methods in clinical medicine are chemotherapy and radiotherapy, with severe side effects. In recent years, some new treatments for cancer have been discovered, such as gene therapy, photothermal therapy and so on. In this paper, we will propose a new method of treatment of cancer, called gel therapy, without the use of anticancer drugs. The high concentration of glutathione (GSH) in tumor cells can be selectively used to induce the death of cancer cells without external stimulation, thus avoiding the side effects on normal tissue cells. In this paper, calcium alginate gel (CaAlg) was transported into cancer cells by bovine serum albumin capsule, and the gel was released under the action of high concentration of GSH in cancer cells, resulting in situ gelation of the cytoplasm and killing of cancer cells. The main works are as follows: (1) the construction of bovine serum albumin capsules: the protein-polymer coupling (BSA-NH _ 2 / PNIPAAm), which was obtained by the reaction of polyN-isopropylacrylamide (PNIPAAm) and amino bovine serum albumin (BSA-NH2), was used as the unit. Protein capsules were prepared by Pickering microemulsion method. The stability of the structure was obtained by adjusting the concentration of the unit, the ultrasonic time and the ultrasonic power. (2) preparation and characterization of protein capsules loaded with sodium alginate (NaAlg). The formation of the gel was proved by SEM TEM, and then the gel was induced by glutathione, and the protein capsule loaded with sodium alginate was observed by microscope. The process of gel formation and gel release. (3) in situ detection of the gelation process of cells by AFM: the Young's modulus of the cells was measured and calculated by AFM, and the life state of the cells was reflected by the changes of the Young's modulus of the cells. The process of gel formation and gel release was further demonstrated. (4) in vitro experiments: firstly, the effects of culture time and capsule concentration on endocytosis were studied. Then the cytotoxicity test (MTT) was performed on human hepatoma cells (HepG2) and mouse embryonic fibroblasts (NIT3T3), and the cell activity was tested by MTT assay. However, there was no significant effect on normal cells, indicating that this method can selectively kill cancer cells.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R73-36;O648.17

【相似文献】