本文選題：RGD + MS2　； 參考：《北京協(xié)和醫(yī)學院》2016年博士論文

【摘要】：阿霉素(Doxorubicin, DOX)作為化療藥物在腫瘤的臨床治療中被廣泛應用。但由于阿霉素水溶性差,組織選擇特異性低,因此,在治療過程中,會對患者造成嚴重的副作用。此外,長期使用阿霉素將會誘導腫瘤細胞產(chǎn)生耐藥性。這些因素限制了阿霉素在腫瘤治療中的進一步應用。納米藥物近年來不斷興起,成為腫瘤治療的一種策略。納米藥物利用增強的藥物滲透及潴留效應進入腫瘤部位發(fā)揮作用。與小分子相比,納米藥物有如下優(yōu)勢：具有納米尺度的化療藥物因其分子量大而在體內(nèi)具有較長的半衰期；此外,納米載體可裝載大量藥物,從而使腫瘤部位的藥物濃度增加；不僅如此,納米藥物具有良好的組織選擇性,因此,對其它器官的副作用較小。MS2病毒樣顆粒是一種無感染性,耐RNA/DNA酶,可在體外大量表達的具有納米尺度的生物材料,因此,可作為納米材料用于藥物遞送。但由于MS2缺乏靶向特異性,需要特異性的配體以提高其靶向特異性。本研究采用MS2作為載體,利用兩功能交聯(lián)劑SM(PEG)24將RGD和化療藥物阿霉素與MS2交聯(lián),制備出具有靶向特異性的納米藥物RGD-MS2-DOX。電鏡實驗和動態(tài)光散射掃描實驗顯示所制備的RGD-MS2-DOX形態(tài)均一,分散性良好,徑粒約為30nm。為觀察RGD-MS2-DOX的腫瘤殺傷功能,我們分別從細胞水平和動物水平兩個方面對該藥物進行研究。在細胞水平,我們采用高表達a vβ3的Hela細胞、HUVECs細胞和HepG2細胞這三種腫瘤細胞進行實驗,發(fā)現(xiàn)RGD-MS2-DOX可通過特異性的配體-受體介導的胞吞作用進入細胞,并且進入細胞的量多于RAD-MS2-DOX和DOX。不僅如此,腫瘤細胞攝取RGD-MS2-DOX是時間依賴性的,隨著藥物作用時間的延長而增加藥物的攝入量。電鏡實驗發(fā)現(xiàn)RGD-MS2-DOX進入細胞后可釋放出DOX,進入細胞核,發(fā)揮腫瘤殺傷功能。與DOX相比,RGD-MS2-DOX對Hela細胞和HepG2細胞的增殖具有較為明顯的抑制效果。為了觀察RGD-MS2-DOX的安全性與體內(nèi)腫瘤殺傷功能,我們選擇長有人肝癌HepG2的Balb/c裸鼠作為腫瘤模型,發(fā)現(xiàn)RGD-MS2-DOX的毒副作用小,并且抑瘤效果明顯。本研究建立了以MS2 VLPs作為納米材料的新型廣譜的主動靶向納米藥物RGD-MS2-DOX,該納米藥物可針對多種類型的腫瘤細胞發(fā)揮靶向殺傷作用,并在體內(nèi)外呈現(xiàn)出良好的抗腫瘤作用,值得進一步深入系統(tǒng)的研究。
[Abstract]:Doxorubicin (DOX) is widely used as a chemotherapeutic agent in the clinical treatment of cancer. However, due to poor water solubility and low tissue selection specificity, adriamycin can cause serious side effects in the course of treatment. In addition, long-term use of adriamycin will induce drug resistance in tumor cells. These factors limit the further application of adriamycin in tumor therapy. Nanopharmaceuticals are emerging in recent years as a strategy for tumor therapy. Nanopharmaceuticals increase drug penetration and retention into the tumor site to play a role. Compared with small molecules, nanopharmaceuticals have the following advantages: nanoscale chemotherapeutic drugs have a longer half-life in vivo due to their high molecular weight, in addition, nano-carriers can load a large number of drugs, thus increasing the concentration of drugs in tumor sites; Not only that, nanopharmaceuticals have good tissue selectivity, so the side effects on other organs are small. MS2 virus-like particles are noninfectious, RNA-resistant, RNA-resistant DNAases, and can be expressed in vitro with nanoscale biomaterials, so, It can be used as nanometer material for drug delivery. However, due to the lack of targeting specificity of MS2, specific ligands are needed to improve its targeting specificity. In this study, RGD-MS2-DOX was prepared by cross-linking RGD and doxorubicin with MS2 by using two functional crosslinking agent SM (PEG) 24. The results of electron microscopy and dynamic light scattering scanning showed that the prepared RGD-MS2-DOX was homogeneous in morphology and good in dispersion. The diameter of the prepared RGD-MS2-DOX was about 30 nm. In order to observe the tumor killing function of RGD-MS2-DOX, we studied the drug from cell level and animal level. At the cell level, we used three kinds of tumor cells, HUVECs and HepG2 cells, which expressed highly a v 尾 3. We found that RGD-MS2-DOX could enter the cells through specific ligand-receptor mediated endocytosis. And the number of entering cells is more than RAD-MS2-DOX and DOX. Moreover, the uptake of RGD-MS2-DOX by tumor cells was time dependent and increased with the prolongation of drug action time. Electron microscopy showed that RGD-MS2-DOX could release DOX into the nucleus and play a role in tumor killing. Compared with DOX, RGD-MS2-DOX could inhibit the proliferation of Hela cells and HepG2 cells. In order to observe the safety of RGD-MS2-DOX and tumor killing function in vivo, Balb / c nude mice with long liver cancer HepG2 were selected as tumor model. The results showed that RGD-MS2-DOX had little toxicity and obvious tumor inhibition effect. In this study, a novel broad-spectrum active target drug RGD-MS2-DOX, using MS2VLPs as nanomaterials, was established. The drug can be targeted at various types of tumor cells and has a good anti-tumor effect in vivo and in vitro. It is worth further and systematic study.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R730.5
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本文編號：2084325