靶向纳米粒子的绿色合成及抑制肿瘤转移研究

发布时间：2018-03-17 08:42

  本文选题：纳米材料　切入点：植物多酚　出处：《暨南大学》2016年博士论文　论文类型：学位论文

【摘要】：肿瘤的转移是癌症发病率和死亡率的主要原因。虽然早期诊断和抗肿瘤药物的研发工作取得了很大进展,使癌症患者的生存率明显提高。然而仍然有许多因素限制了抗肿瘤药物的治疗疗效,复发和转移仍是未解决的难题。肿瘤的浸润和迁移是由多基因参与,多条信号转导途径共同交叉调控的一个复杂过程。因此针对肿瘤侵袭转移中的一些关键环节专门设计有效作用靶点,成为开发抗肿瘤侵袭和转移药物研究的一大热点。为了适合在生物医学领域中应用,理想纳米材料的合成方法是避免使用有毒的化学试剂,并能更好地控制纳米粒子的形状和大小。因此,基于对肿瘤转移分子水平上的认识,本论文从天然产物植物多酚中寻找具有良好活性的植物成分,从形貌、大小和靶向配体的角度设计合成了三种纳米材料,系统地研究了其抗肿瘤侵润和转移的作用,并对其抑制机制进行了深入探讨。本研究论文分为四章:第一章:绪论部分,介绍了肿瘤浸润转移的分子生物学机制。阐明了纳米粒子的绿色合成方法及其物理化学性质对生物系统的影响。最后综述了抗侵袭和转移纳米药物的研究进展。第二章:为了筛选合适形貌的硒纳米粒子进行细胞生物学行为的研究,我们利用鞣酸(TA)作为还原剂和表面修饰剂,制备出了梭型、球型和花型三种不同形貌的TA-Se NPs。相对于梭型和花型,球型TA-Se NPs表现出更好的细胞吸收和抑制癌细胞生长的效果。因此我们选择球型TA-Se NPs(B)做为进一步实验的研究对象。研究发现TA-Se NPs能够抑制体内和体外由b FGF诱导的血管生成。在对纳米粒子抑制血管生成机理研究中发现,TA-Se NP能与b FGF结合,导致b FGF与高亲和力受体(FGFR)的结合力受到影响,从而减少了MAPK/Erk和P13K/AKT通路信号分子的激活,进而抑制了血管生成。同时TA-Se NP也能通过下调细胞内b FGF的表达来参与对内皮细胞的迁移、管形成等的调节。第三章:纳米硒的绿色化学合成往往会导致纳米的尺寸较大,从而限制了其作为抗肿瘤药物的应用。因此我们利用没食子酸(GA)作为还原剂和封端剂,通过简单的绿色合成方法制备出粒径为60 nm的硒/钌复合纳米粒子(GA-Se/Ru NPs)。Se/Ru复合纳米粒子的粒径和形态能通过改变GA的浓度加以调控,并且Ru也是调节和控制尺寸的关键性因素。此外GA-Se/Ru NPs能够通过诱导细胞凋亡来抑制He La细胞的生长,而对正常的HK-2细胞无明显毒性。另外GA-Se/Ru NPs通过下调MMP-2和MMP-9的蛋白表达有效地抑制了宫颈癌He La细胞的浸润和迁移。研究结果表明由GA介导合成的Se/Ru复合纳米粒子展现了更强的抗癌效果。第四章:功能化的纳米粒子可以通过受体介导的内吞作用促进肿瘤细胞对纳米粒子的摄入,从而能更有效地发挥其抗肿瘤的治疗作用。在这一章中,由于表没食子儿茶素没食子酸酯(EGCG)与在肝癌细胞中过表达的67-k Da层粘连蛋白受体(67LR)具有高亲和力。因此我们设计用EGCG功能化负载有发光钌配合物的纳米钌粒子(Ru NPs),以实现更高效的抗肿瘤功效。合成出的纳米粒子(Ru BB-loaded EGCG-Ru NPs)对肝癌细胞具有较高的选择性。细胞对Ru BB-loaded EGCG-Ru NPs的内化作用可以被67LR抗体和层粘连蛋白抑制,表明67LR介导的内吞作用在肝癌细胞对Ru BB-loaded EGCG-Ru NPs的摄入中起到了关键作用。TEM和共聚焦显微图像显示纳米粒子主要聚集在SMMC-7721细胞的细胞质中。进一步的结果表明EGCG功能化的纳米粒子以其特异性分子靶点方式显示出增强的抗肿瘤活性。Ru BB-loaded EGCG-Ru NPs表现出对人肝癌细胞SMMC-7721具有ROS介导的直接细胞毒性,且具有促肿瘤细胞凋亡及抗肿瘤细胞侵袭作用,而对正常细胞L-02无细胞毒性。此外,裸鼠移植瘤体内模型实验表明Ru BB-loaded EGCG-Ru NPs具有较高的抗肿瘤功效。
[Abstract]:Tumor metastasis is a major cause of cancer morbidity and mortality. Although early diagnosis and antitumor drug research and development work has made great progress, the survival rate of cancer patients improved significantly. However, there are still many factors that restrict the therapeutic effect of anticancer drugs, the problem of recurrence and metastasis is still unresolved. Tumor invasion and migration is a multi gene, a complex process of multiple signal transduction regulation. Therefore the common cross tumor invasion of some key links in the transfer of specially designed effective targets, has become a hot topic in the development of tumor invasion and metastasis. In order to study drugs suitable for application in the biomedical field, the ideal synthesis method nano material is to avoid the use of toxic chemical reagents, and can better control the shape and size of nanoparticles. Therefore, based on the molecular level of tumor metastasis Understanding the search of plant ingredients, with good activity from natural products of plant polyphenols from morphology, size and targeting ligand design from the perspective of three kinds of nano materials were synthesized and studied systematically the tumor invasion and metastasis, and the inhibition mechanism were discussed in this paper. Divided into four chapters: the first chapter: the introduction part, introduces the molecular mechanism of tumor invasion and metastasis. Illustrates the influence of green synthesis of nanoparticles and their physical and chemical properties of biological systems. Finally summarizes the research progress of nano drug and anti invasion transfer. The second chapter: in order to study the selenium nanoparticles screening suitable for biological morphology the behavior of cells, we use tannic acid (TA) as the reducing agent and surface modifying agent, was prepared by spindle type, ball type and pattern of three different morphologies of TA-Se NPs. relative to the spindle and flowers Type, ball type TA-Se NPs showed better absorption of cells and inhibiting the growth of cancer cells. So we choose the ball type TA-Se NPs (B) as the research object for further study. The study found that TA-Se NPs can inhibit the angiogenesis induced by B FGF in vivo and in vitro. In the study found to inhibit the generation mechanism of vascular nanoparticles in TA-Se, the combination of NP and B FGF, B FGF and lead to high affinity receptor (FGFR) binding force is affected, thereby reducing the activation of MAPK/Erk and P13K/AKT signaling pathway, thereby inhibiting angiogenesis. TA-Se and NP can down regulate expression of intracellular B FGF to participate in the migration of endothelial cells the tube formation regulation. The third chapter: green chemical synthesis of nano selenium nanoparticles often leads to a larger size, which limits its application as anticancer drugs. So we use gallic acid (GA) as Reductant and capping agent, through the green synthesis method has the advantages of simple preparation of a particle size of 60 nm selenium / ruthenium composite nanoparticles (GA-Se/Ru NPs).Se/Ru composite nanoparticles with particle size and morphology can be regulated by changing the concentration of GA, and Ru is the key factor to regulate and control the size. In addition to GA-Se/Ru NPs to inhibit He La by inducing the apoptosis of cell growth, and has no obvious toxicity to normal HK-2 cells. In addition GA-Se/Ru NPs through down-regulation of MMP-2 and MMP-9 protein expression was effectively inhibited La cell He cancer invasion and migration. The results show that GA mediated by Se/Ru composite nanoparticles synthesized show anticancer effect stronger. The fourth chapter: the endocytosis of nanoparticles functionalized by receptor mediated tumor cells to promote the intake of nanoparticles, which can exert its anti-tumor therapy more effectively . in this chapter, the epigallocatechin gallate (EGCG) and the expression of 67-k in HCC cells Da laminin receptor (67LR) with high affinity. So we design using EGCG functionalized nano luminescent ruthenium ruthenium complexes (Ru NPs), in order to achieve more efficient anti-tumor efficacy. Nanoparticles synthesized (Ru BB-loaded EGCG-Ru NPs) has a high selectivity to hepatocellular carcinoma cells. Ru BB-loaded EGCG-Ru cells on the internalization of NPs can be 67LR antibody and laminin inhibition showed that 67LR mediated endocytosis plays a key role in.TEM and confocal micrograph of nanoparticles mainly accumulated in the cytoplasm of SMMC-7721 cells in human hepatocellular carcinoma cells Ru BB-loaded EGCG-Ru intake on NPs. Further results show that EGCG nanoparticles functionalized with specific molecular targets of Shi Xian Is shown to enhance the antitumor activity of.Ru BB-loaded EGCG-Ru NPs has shown direct cytotoxicity mediated by ROS on human hepatocellular carcinoma cell line SMMC-7721, and has the effect of promoting tumor cell apoptosis and invasion of tumor cells, but no cytotoxicity to normal cells L-02. In addition, nude mice model in vivo experiments show that Ru BB-loaded EGCG-Ru NPs has high the anti tumor effect.

【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R73-37
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本文编号：1623990