超顺磁性葡聚糖氧化铁纳米颗粒的制备及其作为基因载体的可行性研究

目的磁性纳米颗粒作为基因载体在肿瘤基因治疗中的应用得到了迅速发展。为了能获得驱动目的基因高效稳定表达、安全无害、靶向性高、简便的新型非病毒型基因导入和治疗系统,本研究探讨超顺磁性葡聚糖氧化铁纳米颗粒(superparamagnetic dextran i- ron oxide nanoparticles,SDION)的制备及其作为体外基因载体的可行性。方法采用化学共沉淀法制作SDION,通过丙烯葡聚糖凝胶S-300HR色谱和离心法分离sDl0N,用透射电镜、粒度分析仪和磁力计对SDION进行分析。以绿色荧光蛋白(GFP-C2)质粒为靶基因,通过氧化还原法构建SDION-GFP-C2复合物,用紫外分光光度计和琼脂糖凝胶电泳检测两者的结合率。以脂质体转染作为对照,荧光显微镜分别观察SDION和脂质体体外转染GFP-C2入膀胱癌细胞BIU-87的转染效率。结果SDION直径在3-8 nm之间,有效粒径为59.2 nm,比饱和磁化强度为0.23 emtb,g。分别经10 mmol/L的高碘酸钠氧化、0.5 moL/L的硼氢化钠还原作用后的SDION和GFP的结合比例最大,SDUIN对GFP DNA的转染效率为45%左右,明显高于脂质体的转染效率(30%左右)。结论SIION可通过氧化还原反应与GFP质粒相连,在体外可将GFP基因成功转染入人膀胱癌BIU-87细胞。     

超顺磁性葡聚糖氧化铁纳米颗粒的制备及其作为基因载体的可行性研究

背景与目的：磁性纳米颗粒作为基因载体在肿瘤基因治疗中的应用得到了迅速发展。为了能获得驱动目的基因高效稳定表达、安全无害、靶向性高、简便的新型非病毒型基因导入和治疗系统,本研究探讨超顺磁性葡聚糖氧化铁纳米颗粒(superparamapnetic dextran iron oxide nanoparticles,SDION)的制备及其作为体外基因载体的可行性.方法：采用化学共沉淀法制作SDION,通过丙烯葡聚糖凝胶S-300HR色谱和离心法分离SDION,用透射电镜、粒度分析仪和磁力计对SDION进行分析。以绿色荧光蛋白(GFP-C2)质粒为靶基因,通过氧化还原法构建SDION-GFP-C2复合物,用紫外分光光度计和琼脂糖凝胶电泳检测两者的结合率。以脂质体转染作为对照,荧光显微镜分别观察SDION和脂质体体外转染GFP-C2入膀胱癌细胞BIU-87的转染效率。结果：SDION直径在3～8nm之间,有效粒径为59．2nm,比饱和磁化强度为0．23emu／g。分别经10mmol／L的高碘酸钠氧化、0．5mol／L的硼氢化钠还原作用后的sDION和GFP的结合比例最大,sDION对GFPDNA的转染效率为45％左右,明显高于脂质体的转染效率(30％左右)。结论：SDION可通过氧化还原反应与GFP质粒相连,在体外可将GFP基因成功转染入人膀胱癌BIU-87细胞。     

超顺磁性葡聚糖氧化铁纳米颗粒的制备及其作为基因载体的可行性研究(英文)

目的磁性纳米颗粒作为基因载体在肿瘤基因治疗中的应用得到了迅速发展。为了能获得驱动目的基因高效稳定表达、安全无害、靶向性高、简便的新型非病毒型基因导入和治疗系统,本研究探讨超顺磁性葡聚糖氧化铁纳米颗粒(superparamagnetic dextran i-ron oxide nanoparticles,SDION)的制备及其作为体外基因载体的可行性。方法采用化学共沉淀法制作 SDION,通过丙烯葡聚糖凝胶 S-300HR 色谱和离心法分离 SDION,用透射电镜、粒度分析仪和磁力计对 SDION 进行分析。以绿色荧光蛋白(GFP-C2)质粒为靶基因,通过氧化还原法构建 SDION-GFP-C2复合物,用紫外分光光度计和琼脂糖凝胶电泳检测两者的结合率。以脂质体转染作为对照,荧光显微镜分别观察 SDION 和脂质体体外转染 GFP-C2入膀胱癌细胞 BIU-87的转染效率。结果 SDION 直径在3～8nm 之间,有效粒径为59.2nm,比饱和磁化强度为0.23 emu/g。分别经10 mmol/L 的高碘酸钠氧化、0.5mol/L 的硼氢化钠还原作用后的 SDION 和 GFP 的结合比例最大,SDION 对 GFP DNA 的转染效率为45%左右,明显高于脂质体的转染效率(30%左右)。结论 SDION 可通过氧化还原反应与 GFP 质粒相连,在体外可将 GFP 基因成功转染入人膀胱癌 BIU-87细胞。     

氧化铁纳米颗粒作为p53基因载体并转染肺癌细胞的可行性研究

目的:体外评价表面修饰多聚赖氨酸的氧化铁磁性纳米颗粒作为野生型p53基因载体并转染肺癌细胞的可行性。方法:以碱沉淀法制成外包葡聚糖的氧化铁磁性生物纳米颗粒(dextran coated iron oxide nanoparticles,DCIONP),在其表面修饰多聚赖氨酸制成多聚赖氨酸-氧化铁纳米颗粒(dextran coated iron oxide nanoparticles modified with polyL-lysine,pll-DCIONP)。扫描电镜观察其形态特征,用激光粒度检测仪测定其粒度分布和Zeta表面电位;分光光度计及琼脂糖凝胶电泳分析pll-DCIONP与野生型p53基因的结合力及其复合物抵抗DNase-Ⅰ和血清消化的能力;pll-DCIONP作为野生型p53基因载体转染人肺腺癌细胞A549/CDDP,RT-PCR以及Western印迹法分析细胞内p53基因的表达。结果:pll-DCIONP的直径在60~80nm之间,Zeta电位为 19.6 mV;pll-DCIONP无论在酸性、中性及碱性的条件下,均可与野生型p53基因结合,二者质量比为1∶1时结合力最强;pll-DCIONP/wt-p53复合物能抵抗DNase-Ⅰ和血清对野生型p53基因的消化作用;以pll-DCIONP作为野生型p53基因载体转染人肺腺癌细胞,随着时间延长,细胞内p53基因含量持续增高,而以脂质体作为转染载体时随着时间延长细胞内p53基因含量递减。结论:pll-DCIONP可作为野生型p53基因理想的转染载体,并能持续高效地将外源性p53基因转染入肺癌细胞中。     

氧化铁磁性纳米颗粒介导wt-p53基因对肺腺癌细胞增殖和凋亡的影响

目的：探讨氧化铁磁性纳米颗粒介导野生型p53基因（wild type p53,wt-p53）对耐顺铂人肺腺癌细胞A549/DDP增殖抑制和凋亡诱导的作用。方法：氧化铁磁性纳米颗粒介导wt-p53转染肺腺癌细胞A549/DDP作为实验组,以纳米颗粒介导空载体pcDNA3转染作为阴性对照组,脂质体介导wt-p53转染作阳性对照组。MTT法和绘制生长曲线观察基因转染对A549/DDP细胞增殖抑制作用,荧光显微镜、流式细胞术观察其对A549/DDP细胞诱导凋亡作用,RT-PCR检测其对A549/DDP细胞Bax mRNA表达的影响。结果：氧化铁磁性纳米颗粒介导wt-p53对人肺腺癌细胞A549/DDP增殖有持续的抑制作用,而以脂质体介导wt-p53对增殖抑制作用持续时间短暂;纳米颗粒介导wt-p53对人肺腺癌细胞A549/DDP诱导凋亡作用明显强于以脂质体载体;同时介导wt-p53上调Bax mRNA表达水平的作用也明显强于以脂质体载体。结论：氧化铁磁性纳米颗粒介导wt-p53转染对人肺腺癌细胞A549/DDP有持续的增殖抑制和诱导凋亡的作用。     

磁性纳米颗粒介导基因治疗乳腺癌实验研究

 目的 探讨新型基因载体PEI包裹的磁性纳米颗粒polyMAG-1000介导TRAIL基因治疗乳腺癌的可行性，观察TRAIL基因转染乳腺癌细胞后的治疗作用。方法 以polyMAG-1000为基因载体，联结TRAIL质粒DNA后转染人乳腺癌细胞株MCF-7细胞，用Tunel法检测细胞的凋亡，用流式细胞仪检测细胞的凋亡率，以空载体作为阴性对照；脂质体转染TRAIL基因作阳性对照。结果 Tunel法可见MCF-7细胞经polyMAG1000和脂质体转染TRAIL基因后均可见发生凋亡的细胞，流式细胞仪检测polyMAG-1000转染的细胞凋亡率为25．11％±2．85％，脂质体转染的细胞凋亡率为18．31％±2．44％（P〈0．05）。结论 TRAlL对乳腺癌细胞MCF-7具有凋亡效应，PEI包裹的磁性纳米颗粒介导的TRAIL基因治疗在肿瘤的基因治疗中具有应用前景。     

多聚赖氨酸-硅纳米颗粒的生物相容性研究

背景与目的：多聚赖氨酸-硅纳米颗粒是一种新型的非病毒纳米颗粒基因传递载体。本研究主要阐述其生物相容性,为其进一步的体内应用提供实验依据。方法：细胞转染和流式细胞仪分析在含血清培养基中多聚赖氨酸-硅纳米颗粒介导质粒DNA和反义寡核苷酸的转染能力,随后通过血浆蛋白反应滤过试验和红细胞聚集试验进一步评价其生物相容性。结果：在含血清培养基中,多聚赖氨酸-硅纳米颗粒介导质粒DNA和反义寡核苷酸传递的能力显著下降。多聚赖氨酸-硅纳米颗粒／DNA(／ODN)复合物可与小鼠血浆蛋白成分结合,并可引起红细胞聚集。结论：多聚赖氨酸-硅纳米颗粒可能与含血清培养基中的血浆蛋白相互作用,影响体外细胞转染效率。多聚赖氨酸-硅纳米颗粒可能与体内外周血血浆蛋白和外周血红细胞反应而影响其体内基因传递能力,其生物相容性还有待于进一步提高。     

氧化铁磁性纳米颗粒介导wt p53基因对肺腺癌细胞增殖和凋亡的影响

目的: 探讨氧化铁磁性纳米颗粒介导野生型p53基因（wild type p53,wtp53）对耐顺铂人肺腺癌细胞A549/DDP增殖抑制和凋亡诱导的作用。 方法:氧化铁磁性纳米颗粒介导wtp53转染肺腺癌细胞A549/DDP作为实验组，以纳米颗粒介导空载体pcDNA3转染作为阴性对照     

HER2/neu胞外区片段基因免疫诱导特异细胞免疫及其抗瘤效应研究 *

目的：了解鱼精蛋白应用于血管内皮生长因子受体介导的靶向性非病毒载体的可行性。方法：ＣＶ１,ＣＶ２靶向性非病毒载体来比较多聚赖氨酸与鱼精蛋白对靶向性基因转移复合体携带ＤＮＡ的能力及体外基因转移效率的影响。结果：在Ａ３７５细胞中,鱼业 白与多聚赖氨酸参与形成的复合基因导入率都为５０％左右。在ＡＢＡＥ细胞中,鱼精蛋白参与形成的复合体基因导入率只有２０％左右,而多聚赖酸可达７０％左右。鱼精蛋白参与形成的复     

LHRH-conjugated Magnetic Iron Oxide Nanoparticles for Detection of Breast Cancer Metastases

Summary Targeted delivery of superparamagnetic iron oxide nanoparticles (SPIONs) could facilitate their accumulation in metastatic cancer cells in peripheral tissues, lymph nodes and bones and enhance the sensitivity of magnetic resonance imaging (MRI). The specificities of luteinizing hormone releasing hormone (LHRH) and luteinizing hormone/chorionic gonadotropin (LH/CG)- bound SPIONs were tested in human breast cancer cells in vitro and were found to be dependent on the receptor expression of the target cells, the time of incubation and showed saturation kinetics. In incubations with MDA-MB-435S.luc cells, the highest iron accumulation was 452.6 pg Fe/cell with LHRH-SPIONs, 203.6 pg Fe/cell with β-CG-SPIONs and 51.3 pg Fe/cell with SPIONs. Incubations at 4 °C resulted in 1.1 pg Fe/cell. Co-incubation with the same ligands (βCG or LHRH) decreased the iron accumulation in each case. LHRH-SPIONs were poorly incorporated by macrophages. Tumors and metastatic cells from breast cancer xenografts were targeted in vivo in a nude mouse model. LHRH-SPION specifically accumulated in cells of human breast cancer xenografts. The amount of LHRH-SPION in the lungs was directly dependent on the number of metastatic cells and amounted to 77.8 pg Fe/metastastic cell. In contrast, unconjugated SPIONs accumulated in the liver, showed poor affinity to the tumor, and were not detectable in metastatic lesions in the lungs. LHRH-SPION accumulated in the cytosolic compartment of the target cells and formed clusters. LHRH-SPIONs did not accumulate in livers of normal mice. In conclusion, LHRH conjugated SPIONs may serve as a contrast agent for MR imaging in vivo and increase the sensitivity for the detection of metastases and disseminated cells in lymph nodes, bones and peripheral organs.     

钆-叶酸-四氧化钴铁磁性纳米粒子对卵巢癌细胞靶向性能的影响

[目的]探讨钆-叶酸-四氧化钴铁磁性纳米粒子通过靶向结合卵巢癌COC1细胞,在核磁共振成像时对卵巢癌组织对比度的性能影响.[方法]利用透射电镜观察纳米粒子靶向进入卵巢癌细胞的过程,采用荧光光谱法测定不同浓度下纳米粒子进入细胞的定量,MTT法检测纳米粒子的细胞毒性,再通过核磁共振成像技术分析不同浓度纳米粒子对T1层面成像、T2层面成像的作用.[结果]纳米粒子能够靶向进入卵巢癌COC1细胞,并呈剂量依赖性;在进入细胞48h内未显示出细胞毒性,并能够在T1、T2两个层面增加核磁共振成像的组织对比度.[结论]钆-叶酸-四氧化钴铁磁性纳米粒子能够增加卵巢癌核磁共振成像组织的对比度,提高其分辨率,有可能成为一种既能改变T1成像,又能改变T2成像分辨率的新的靶向磁性造影剂.     

Recombinant Interleukin-1 Receptor Antagonist Conjugated to Superparamagnetic Iron Oxide Nanoparticles for Theranostic Targeting of Experimental Glioblastoma

Cerebral edema commonly accompanies brain tumors and contributes to neurologic symptoms. The role of the interleukin-1 receptor antagonist conjugated to superparamagnetic iron oxide nanoparticles (SPION–IL-1Ra) was assessed to analyze its anti-edemal effect and its possible application as a negative contrast enhancing agent for magnetic resonance imaging (MRI). Rats with intracranial C6 glioma were intravenously administered at various concentrations of IL-1Ra or SPION–IL-1Ra. Brain peritumoral edema following treatment with receptor antagonist was assessed with high-field MRI. IL-1Ra administered at later stages of tumor progression significantly reduced peritumoral edema (as measured by MRI) and prolonged two-fold the life span of comorbid animals in a dose-dependent manner in comparison to control and corticosteroid-treated animals (P < .001). Synthesized SPION–IL-1Ra conjugates had the properties of negative contrast agent with high coefficients of relaxation efficiency. In vitro studies of SPION–IL-1Ra nanoparticles demonstrated high intracellular incorporation and absence of toxic influence on C6 cells and lymphocyte viability and proliferation. Retention of the nanoparticles in the tumor resulted in enhanced hypotensive T₂-weighted images of glioma, proving the application of the conjugates as negative magnetic resonance contrast agents. Moreover, nanoparticles reduced the peritumoral edema confirming the therapeutic potency of synthesized conjugates. SPION–IL-1Ra nanoparticles have an anti-edemal effect when administered through a clinically relevant route in animals with glioma. The SPION–IL-1Ra could be a candidate for theranostic approach in neuro-oncology both for diagnosis of brain tumors and management of peritumoral edema.     

磁流体热疗对小鼠胰腺癌治疗作用的研究

目的:探讨磁流体热疗对小鼠胰腺癌的治疗作用.方法:选择4周龄雌性昆明种小鼠,利用我们前期建株的小鼠胰腺癌细胞系MPC-83建立小鼠胰腺癌皮下肿瘤模型,将0.2mL浓度为1.2g/mL磁流体直接注射到肿瘤内,24h后置入交变磁场中升温至46℃和50℃,作用30min,观察磁流体热疗对荷瘤小鼠的影响及其病理学检查.结果:实验组的肿瘤瘤体中心温度分别迅速升温并稳定在46℃和50℃,而直肠温度保持在30℃～36℃之间.热疗后即刻组织病理学见肿瘤细胞呈凋亡、坏死样改变.热疗后14相似文献   

Targeting Glioblastoma with the Use of Phytocompounds and Nanoparticles

Glioblastoma multiforme (GBM) are extremely lethal and still poorly treated primary brain tumors, characterized by the presence of highly tumorigenic cancer stem cell (CSC) subpopulations, considered responsible for tumor relapse. In order to successfully eradicate GBM growth and recurrence, new anti-cancer strategies selectively targeting CSCs should be designed. CSCs might be eradicated by targeting some of their cell surface markers and transporters, inducing their differentiation, impacting their hyper-glycolytic metabolism, inhibiting CSC-related signaling pathways and/or by targeting their microenvironmental niche. In this regard, phytocompounds such as curcumin, isothiocyanates, resveratrol and epigallocatechin-3-gallate have been shown to prevent or reverse cancer-related epigenetic dysfunctions, reducing tumorigenesis, preventing metastasis and/or increasing chemotherapy and radiotherapy efficacy. However, the actual bioavailability and metabolic processing of phytocompounds is generally unknown, and the presence of the blood brain barrier often represents a limitation to glioma treatments. Nowadays, nanoparticles (NPs) can be loaded with therapeutic compounds such as phytochemicals, improving their bioavailability and their targeted delivery within the GBM tumor bulk. Moreover, NPs can be designed to increase their tropism and specificity toward CSCs by conjugating their surface with antibodies specific for CSC antigens, with ligands or with glucose analogues. Here we discuss the use of phytochemicals as anti-glioma agents and the applicability of phytochemical-loaded NPs as drug delivery systems to target GBM. Additionally, we provide some examples on how NPs can be specifically formulated to improve CSC targeting. Open image in new window