本文選題：金納米材料 + 腫瘤治療　； 參考：《東南大學》2017年博士論文

【摘要】：金納米顆粒是一種非常重要的金屬納米材料,其獨特的光學性質,易控的表面化學性能和良好的生物相容性使其成為基礎研究及應用研究的熱點。由于金具有較高的原子序數(shù),可以增強放射線對腫瘤區(qū)域的影響,因而金納米材料被廣泛地用于腫瘤放療增敏。某些金納米材料(如金納米棒和金納米刺)的近紅外吸收特性使其具有獨特的光熱效應,從而被廣泛地用于腫瘤的聯(lián)合治療。另外,放射治療還可以誘導細胞自噬應激反應,并且自噬與腫瘤存在雙重關系,這一特性也為腫瘤防治提供了兩種截然不同的思路:抑制自噬提高抗癌治療效果,或激活自噬誘導腫瘤細胞發(fā)生自噬性死亡。因此,目前尚不能盲目地將自噬誘導劑或抑制劑應用于臨床,否則不但不能阻止腫瘤的發(fā)生,反而會促進腫瘤的發(fā)展�；谝陨涎芯勘尘�,本文將以表面粗糙且呈刺突狀的金納米刺(GNSs)為研究主體,圍繞金納米刺在腫瘤放療和熱療中的應用做了一系列研究。具體說來,主要包含以下幾方面的內容:1.比較研究不同形貌的金納米材料的放療增敏特性為了深入闡述不同形貌的金納米材料對放射治療的影響,我們選用了粒徑相同(50 nm左右)且表面均為聚乙二醇(PEG)修飾的三種金納米結構,即金納米球、金納米刺、金納米棒,并對其放療增敏特性進行了系統(tǒng)性地評價。實驗發(fā)現(xiàn),金納米球、金納米刺、金納米棒的放療增敏比(SER)依次為1.62、1.37、1.21,表明三種金納米結構均具有放療增敏效應,且材料的SER與細胞攝取率之間存在正相關性。通過進一步對SER與細胞攝取率的比值進行歸一化分析,結果發(fā)現(xiàn),平均每個金原子對放療增敏的貢獻幾乎一致。也就是說,為實現(xiàn)更大程度的癌細胞殺傷效率,只需要增加金原子的細胞攝取效率即可。通過設計特定形貌的金納米結構可實現(xiàn)最大限度的細胞內化。該項研究對于如何設計有效的金納米結構以用于癌癥的放射治療具有重要的指導意義。2.配體修飾的金納米刺放療增敏作用及細胞自噬應激反應由于金納米刺(GNSs)細胞攝取率低,極大地限制了其放療增敏效果。為此我們制備得到了表面均為PEG修飾且末端帶有不同配體(包括甲氧基、氨基NH2、葉酸FA和細胞穿膜肽TAT)的金納米刺(即GNSs、NH2-GNSs、FA-GNSs和TAT-GNSs),并評價了它們在細胞攝取率和放療增敏效果方面的差異。實驗發(fā)現(xiàn),不同配體修飾的金納米刺的細胞攝取率依次為:TAT-GNSs FA-GNSs NH2-GNSs GNSs。這些材料的放療增敏比SER(依次為2.30、1.84、1.57和1.34)與其細胞攝取率相一致:即TAT-GNSs FA-GNSs NH2-GNSsGNSs。相比于單一 PEG化的金納米刺(端基為甲氧基的GNSs),配體修飾的金納米刺可顯著提高癌細胞的放療增敏效應。我們還進一步從材料的細胞攝取路徑、活性氧產生、線粒體膜電位變化、細胞周期和凋亡等生物學效應角度,深入闡述了放療增敏的潛在機制。結果表明,配體修飾的金納米刺聯(lián)合放療可誘導細胞內活性氧的產生、線粒體膜電位的下降和細胞周期的再分布等。其中,活性氧增多在其中起關鍵作用。此外,我們還探索了細胞自噬與放療增敏之間的關系。結果顯示,自噬作為一種溶酶體依賴的降解途徑,在金納米刺介導的放射治療中扮演重要角色。蛋白印跡檢測結果表明,配體修飾的金納米刺能夠誘導自噬相關蛋白(LC3-Ⅱ)和凋亡相關蛋白(caspase-3)的表達增多;通過對自噬降解底物p62蛋白的定量分析,我們發(fā)現(xiàn)金納米刺可誘導p62的表達增多,這表明金納米刺可損害自噬溶酶體降解能力并導致自噬小體的積累。在自噬抑制劑3-MA的作用下,PEG化的金納米刺及其聯(lián)合放射治療均使細胞凋亡比率明顯增加,即自噬在放療中起保護性作用,而抑制自噬則可極大地促進細胞凋亡。該研究加深了我們對金納米材料細胞攝取規(guī)律的認識,并明確了自噬抑制劑可以顯著增加放療抗癌效果。3.金納米刺介導的高熱效應在光熱和放療協(xié)同抗癌中的應用基于金納米刺良好的光熱效應和放療增敏特性,我們建立了一種基于金納米刺的光熱/放療聯(lián)合抗癌策略。通過大量的體外和體內實驗,系統(tǒng)性地評價了金納米刺介導的光熱/放射聯(lián)合治療效果。金納米刺的SER為1.38,而金納米刺聯(lián)合光熱治療的SER增加到1.63,這表明金納米刺介導的高熱效應顯著增強了放射線對乏氧癌細胞的殺傷效應。此外,體外實驗發(fā)現(xiàn)金納米刺介導的光熱治療、放射治療以及聯(lián)合治療的癌細胞存活率分別為89%、51%和33%。體內實驗顯示金納米刺介導的放射治療、光熱治療以及聯(lián)合治療對腫瘤增殖的抑制率分別為29.8%、70.5%和92.2%。該項研究結果表明,金納米刺介導的熱療和放療具有優(yōu)異的協(xié)同抗癌效果。該研究為多功能金納米材料的開發(fā)和放射治療提供了新的思路。
[Abstract]:Gold nanoparticles are one of the most important metal nanomaterials. Their unique optical properties, easy to control surface chemical properties and good biocompatibility make it a hot spot in basic research and application research. Because of the high atomic number of the metal tools, it can enhance the effect of radiation on the tumor area. Therefore, gold nanoparticles are widely used. The near infrared absorption properties of some gold nanomaterials (such as gold nanorods and gold nanoscale spines) have unique photothermal effects and are widely used in combined treatment of tumors. In addition, radiation therapy can also induce autophagy to induce autophagy and the dual relationship between autophagy and tumor. Two different ideas are provided for the prevention and treatment of tumor: inhibiting autophagy to improve the effect of anticancer therapy, or activating autophagy to induce autophagic cells to occur autophagic death. Therefore, it is not possible to apply autophagy inducers or inhibitors to the clinic blindly. Otherwise, it can not only prevent the occurrence of tumor, but also promote the development of tumor. In this research background, this paper will take a series of studies on the application of gold nano spines (GNSs) with rough surface and stings as the main body. A series of studies have been done around the application of gold nano spines in tumor radiotherapy and hyperthermia. In particular, the main contents are as follows: 1. the radiation sensitization characteristics of gold nanoparticles with different morphologies are compared. The effects of gold nanomaterials with different morphologies were discussed. We selected three gold nanostructures with the same particle size (about 50 nm) and the surface of the gold nanoparticles modified by polyethylene glycol (PEG), namely gold nanoparticles, gold nanoscale and gold nanorods. The radiosensitization ratio (SER) of the gold nanorods is 1.62,1.37,1.21 in turn, indicating that the three gold nanostructures have the effect of radiation sensitization and there is a positive correlation between the material's SER and the cell uptake. The result of the further analysis of the ratio of SER to the cell uptake results shows that the average gold atom is the tribute to the radiation sensitization. The contribution is almost identical. That is to say, in order to achieve greater cancer cell killing efficiency, it only needs to increase the cell uptake efficiency of gold atoms. The maximum cell internalization can be achieved through the design of a gold nanostructure with specific morphologies. This study is heavy on how to design an effective nanoscale structure for cancer radiation therapy. The guiding significance of.2. ligand modified gold nanoscale radiosensitization and cellular autophagy response due to the low uptake rate of gold nanoparticle (GNSs) cells greatly restricts the effect of radiation sensitization. Therefore, we have prepared PEG modification on the surface and the end with different complexes (including methoxy, amino NH2, folic acid FA and cell wear. Gold nanoscale spines (GNSs, NH2-GNSs, FA-GNSs and TAT-GNSs) of membrane peptide TAT, and their differences in cell uptake and radiation sensitization were evaluated. The experimental results showed that the uptake rate of gold nanoparticles modified by different ligands was, in turn, TAT-GNSs FA-GNSs NH2-GNSs GNSs. (TAT-GNSs FA-GNSs NH2-GNSs GNSs.) and SER (2.30,1.84,1.57). And 1.34) in accordance with its cell uptake rate: TAT-GNSs FA-GNSs NH2-GNSsGNSs. is compared to a single PEG gold nanoparticle (terminal methoxy GNSs), and ligand modified gold nanoparticle can significantly increase the radiation sensitization effect of cancer cells. We further from the cell uptake pathway, reactive oxygen generation, and mitochondrial membrane potential change The potential mechanisms of cell cycle and apoptosis, such as cell cycle and apoptosis, are discussed in detail. The results show that ligand modified gold nanoparticle combined with radiotherapy can induce intracellular reactive oxygen species, the decrease of mitochondrial membrane potential and the redistribution of cell cycle. Among them, the increase of reactive oxygen species plays a key role. The relationship between autophagy and radiation sensitization is also explored. The results show that autophagy, as a lysosome dependent degradation pathway, plays an important role in gold nanoparticle mediated radiation therapy. The results of Western blot detection showed that ligand modified gold nanoparticles could induce LC3- II and apoptosis related proteins (CASPA The expression of se-3 is increased. By quantitative analysis of the autophagic degradation substrate p62 protein, we found that gold nanoparticles can induce the increase in the expression of p62, which indicates that gold nanoparticles can damage the degradation ability of autophagosomes and lead to the accumulation of autophagic bodies. Under the action of autophagic inhibitor 3-MA, the PEG nanoscale prickle and its combined radiation therapy are all made The rate of apoptosis is obviously increased, that is, autophagy plays a protective role in radiotherapy, while inhibition of autophagy can greatly promote cell apoptosis. This study deepened our understanding of the uptake of gold nanomaterials. It is clear that autophagy inhibitors can significantly increase the anticancer effect of radiotherapy.3. gold nanoparticle mediated hyperthermia effect in light and heat. Based on the good photothermal effect and radiation sensitization of gold nanoparticles, we established a combination of photothermal / radiotherapy based on gold nanoparticle. Through a large number of in vitro and in vivo experiments, we systematically evaluated the effect of the combined photothermal / radiation therapy on gold nanoparticles. SER For 1.38, the SER of gold nano spines combined with photothermal treatment increased to 1.63, which showed that the high heat effect mediated by gold nano spines significantly enhanced the killing effect of radiation on hypoxic cancer cells. In addition, in vitro, the survival rate of gold nanoparticles mediated photothermal therapy, radiation therapy and combined treatment were 89%, 51% and 33%., respectively. The internal experiment showed that gold nanoparticle mediated radiation therapy, photothermal therapy and combined treatment of tumor proliferation inhibition rate were 29.8%, 70.5% and 92.2%., respectively. The results showed that gold nanoparticle mediated thermotherapy and radiotherapy have excellent synergistic anticancer effects. This study provides the development of multi-functional gold nanomaterials and radiation therapy. A new idea.
【學位授予單位】：東南大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R730.5;TB383.1

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本文編號：1904955