载miR-122的石墨烯-金纳米新型材料对耐药肝癌细胞的靶向治疗研究

In recent years, the nano drugs have been attracted much attention because they offer tumor inhibiting effects due to the unique mode of action. The nano drugs have the following properties: 1, high drug-loaded rate; 2, drug target; 3, drug controlled release; 4, low toxicity. Therefore, it is necessary to explore nano drugs research for development of modern medicine. In the previous report, we have explored the possibility of applying the functionalized drug loaded graphene-gold nanocomposites inducing cancer cells apoptosis, and investigated the relevant mechanism. Our further research, we intend to develop combination of monoclonal P-glycoprotein (P-gp) antibodies and miR-122 loaded on graphene gold nanocomposites (GGMPN), which promoting drug resistance cancer cells apoptosis with drug target and drug controlled release properties. Due to function of P-gp antibody, GGMPN realize targeted drug resistance cancer cells, and combined with glutathione (GSH) which can replace miR-122, realize the function of drug controlled release. GGMPN is used to induce drug resistance HepG2 cells apoptosis. Meanwhile, we investigate related apoptosis protein and apoptosis signal pathway by GGMPN treatment. The different apoptosis mechanism between miR-122 and synergistic effects of GGMPN also should be explored to provide the theoretical basis for the nano drugs therapeutic research. We also demonstrate photothermal therapy to kill cancer cells by using GGMPN with semiconductor laser. Moreover, we investigate inhibition of tumor growth and induce apoptosis effect by using the xenograft tumors nude model. So the present study will provide avenue for the therapeutic approaches to cancer treatments.

近年来纳米药物在癌症治疗中的独特作用备受关注。目前待开发纳米药物应具备以下性质：1.高的载药量，2.靶向性，3.药物控释，4.低毒性。前期试验中，我们开发了靶向促癌凋亡的载柔红霉素石墨烯-金纳米复合物。本研究中，我们拟开发具有靶向控释给药和促耐药癌细胞凋亡作用的负载miR-122的石墨烯-金纳米复合物（GGMPN)，由于GGMPN带有P-糖蛋白抗体可实现对耐药细胞靶向给药，并结合谷胱甘肽（GSH）可以置换miR-122的功能来实现药物控释。GGMPN被用于靶向诱导耐药性的人类HepG2肝癌细胞凋亡。同时对GGMPN促凋亡相关机理进行深入研究，找到促凋亡的信号途径，并和miR-122促凋亡的机理进行比较，为以后开展纳米治疗研究提供理论基础。此外GGMPN还有光热疗作用，本实验拟进一步研究动物实验，并对GGMPN能否有效的促裸鼠肿瘤凋亡作用进行验证，将为高效多功能治疗癌症提供应用前景。

背景：纳米药物因其在肿瘤抑制效果上独特的作用方式，日益受到关注。因此，我们先前探索了能诱导癌细胞凋亡的功能化的、装载药物的石墨烯-金纳米复合材料。.主要研究内容：金纳米颗粒和miR-122形成复合材料后，该复合材料黏附在石墨烯上，影响耐药癌细胞（金颗粒可能被纳入石墨烯）。化疗是肝癌的一种重要治疗方法。由于细胞的多药耐药性，大多数肝癌患者往往忍受了无效的化疗或者从化疗有效逐渐转为化疗无效。当肿瘤细胞暴露于一定的抗癌药物并发生耐药性时，多元耐药性就会产生；它们和其他结构和功能不同的药物有交叉耐药性。多药耐药性的形成机制非常复杂，细胞膜上P-gp表达增加、谷胱甘肽转移酶活性正经是两根重要因素。这种跨学科研究的目的是探讨如何利用miR-122、P-gp抗体和叶酸来做到肿瘤靶向、肿瘤成像和光热治疗。.结果：在目前的研究中，我们开发了一种结合了单克隆P-gp抗体、叶酸和金纳米颗粒装载的miR122的石墨烯复合材料（GGMPN），它能促进耐药HepG2细胞凋亡药物的靶向释放，并能够控制药物释放性能。我们还研究了在体外和体内GGMPN治疗时凋亡相关蛋白和凋亡信号通路。此外，我们通过异种移植肿瘤模型的半导体激光进一步证明了GGMPN的肿瘤生长抑制效应和凋亡诱导效应.结论：总之，我们的研究结果共同表明GGMPN可以作为新的治疗方法控制肿瘤细胞凋亡和增长。