功能化氧化石墨烯运载Stat3-siRNA质粒治疗小鼠恶性黑色素瘤实验研究
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摘要
背景：恶性黑色素瘤是一种常见的恶性肿瘤。早期恶性黑色素瘤通过手术切除，但近年其发病率有明显上升趋势。恶性黑色素瘤具有恶性程度高，转移发生早，累及多个脏器，预后较差，易复发，死亡率高等特点。目前，大多数患者对化疗、放疗均不敏感，生存期短。随着人们对肿瘤分子生物学机制深入研究，基因治疗肿瘤的方法取得了很大进步。利用RNA干涉(RNAinterference，RNAi)技术，有望特异高效地抑制目的基因表达，诱导肿瘤细胞发生凋亡，达到治疗肿瘤的目的。然而如何将目的基因安全、高效的运载到肿瘤组织内，是实现这一目的的瓶颈，近几年，纳米医学的迅速发展不仅为基因载体的研究提供新手段，而且为恶性肿瘤的诊断和治疗带来新的契机。因此，迫切需要开发更具有优异性能的纳米基因传递系统，更好地杀伤肿瘤细胞。
近几年，一种极具应用前景和优异性能的新型纳米材料—石墨烯，以其独特的力学、电学、热学等理化性质引起了研究者极大的兴趣。功能化氧化石墨烯（Graphene Oxide, GO）被生物医学领域的研究者广泛应用。迄今，随着纳米技术的高速发展，越来越多的实验证明，经过表面修饰的GO可以提高基因的转染效率，在生理环境下稳定性好。聚乙烯亚胺（polythyleneimine, PEI）是目前应用较多的阳离子基因传递载体，在生理环境下表面带正电荷，与基因的结合能力强，保护基因免受核酸酶的降解，具有从内涵体逃逸的能力。聚乙二醇（PolyethyleneGlycol，PEG）修饰的氧化石墨烯可以提高GO-PEI稳定性和溶解性，延长复合物在体内的循环时间。所以本实验将采用GO-PEI-PEG反应形成的共聚物作为Stat3siRNA质粒的运载体。
在信号转导通路中，信号转导子和转录激活子3（Signal transducer andactivator of transcription，Stat3）与肿瘤的发生发展密切相关。研究表明，在前列腺癌、肝癌及恶性黑色素瘤等多种肿瘤，Stat3呈现过度表达。Stat3的异常表达通过促进肿瘤新生血管的形成、抑制肿瘤细胞凋亡、促进肿瘤细胞增殖，参与肿瘤的发生过程。因此阻断肿瘤细胞中Stat3通路，可以达到治疗肿瘤的目的。在实验室前期工作中，以RNA干涉技术原理为基础，成功构建Stat3-特异siRNA表达质粒，抑制了前列腺癌、肝癌Stat3基因的过度表达[1,2]，促进肿瘤细胞凋亡。但是，RNAi并不能完全阻断目的基因的表达[3]。光热疗在物理学和生物学具有独特的的优势，与放疗、化疗相比，无毒副作用，具有杀伤肿瘤细胞的作用，可以作为治疗肿瘤的一种辅助手段。因此，光热疗和基因治疗联合策略治疗肿瘤迫在眉睫。
本课题将氧化石墨烯、聚乙烯亚胺、聚乙二醇反应形成共聚物作为肿瘤基因Stat3-特异siRNA质粒的运载体，更好的抑制或杀死肿瘤细胞。同时利用氧化石墨烯吸收红外线的物理学性质，在红外线光照下引起肿瘤组织局部温度升高，从而杀伤肿瘤细胞，进一步探讨功能化氧化石墨烯携带Stat3-特异siRNA质粒对小鼠恶性黑色素瘤的治疗效果及作用机制。
目的：功能化氧化石墨烯携带Stat3-特异siRNA表达质粒体内、体外抑制小鼠恶性黑色素瘤生长及作用机制的探讨。
方法：采用化学法合成氧化石墨烯—聚乙烯亚胺—聚乙二醇（GO-PEI-PEG）基因载体。通过原子力学显微镜、动态光散射等方法，对GO-PEI-PEG粒径、形貌、表面电势等进行表征；研究GO-PEI-PEG在PBS溶液、血清和培养液中的分散性和稳定性。利用琼脂糖凝胶电泳方法，检测在不同质量比条件下，GO-PEI-PEG与si-Stat3结合能力。CCK8法检测GO-PEI-PEG对B16细胞毒性。流式细胞术评价GO-PEI-PEG运载si-Stat3质粒的转染效率。PI染色法检测肿瘤的细胞周期。RT-PCR和Western blot检测Stat3基因及蛋白表达水平；同时检测Stat3相关下游基因的表达情况。建立C57BL/6小鼠恶性黑色素瘤皮下移植瘤模型。透射电子显微镜观察凋亡细胞以及石墨烯的形貌；免疫组织化学方法检测CD34和PCNA的表达；TUNEL试剂盒检测肿瘤细胞凋亡情况；全自动生化分析仪检测各组小鼠血液中BUN、ALT和AST的水平；HE染色观察各组脏器的形态。
结果：成功合成GO-PEI-PEG纳米材料，其表面电势为＋21.02mV，平均粒径为298.9nm，GO-PEI-PEG与si-Stat3质粒结合后形成的复合物，表面电势下降到＋7.52mV，平均粒径增大至339.2nm，在PBS溶液、血清和培养液中具有良好的分散性和稳定性。
体外实验证明，GO-PEI-PEG是一种安全、有效的基因运载体，GO-PEI-PEG携带si-Stat3质粒转染B16细胞，能够观察到绿色荧光蛋白（GFP）的表达。Westernblot和RT-PCR结果显示，GO-PEI-PEG转染si-Stat3组细胞中Stat3基因被干涉，Stat3下游基因表达受到抑制。细胞周期结果显示，GO-PEI-PEG转染si-Stat3组肿瘤细胞周期阻滞在S期，阻止细胞周期的进程。
体内实验证明，GO-PEI-PEG携带Stat3-siRNA质粒联合光照（简称GO-si-Stat3组）显著抑制小鼠恶性黑色素瘤的生长。GO-si-Stat3组的肿瘤瘤重和体积低于其他各组，具有统计学差异。在基因水平和蛋白水平，检测到GO-si-Stat3组Stat3表达受到抑制，Stat3下游调控基因和相关蛋白表达减弱。透射电镜和HE染色观察到凋亡细胞。免疫组织化学结果表明，GO-si-Stat3组增殖的肿瘤细胞减少，微血管生成减少。各组小鼠血液中的BUN、ALT、AST浓度均在正常范围内。HE染色未发现异常的心、肝、脾、肺、肾形态。
结论：功能化氧化石墨烯是一种安全、有效的基因运载体，能够携带Stat3-特异siRNA表达质粒，显著抑制小鼠恶性黑色素瘤的生长。氧化石墨烯具有吸收红外线的物理学性质，联合红外线光照，与Stat3-特异siRNA表达质粒共同发挥抑制小鼠恶性黑色素瘤生长的作用。
Background: Malignant Melanoma is a common cancer, which has high aggressivemalignancy. Patients are effectively treated by surgery at early stages. However, theincidence rates of malignant melanoma in many countries are increasing evidently inrecent years. There are some important biological characteristics in malignantmelanoma, such as high aggressive malignancy, early metastasis, poor prognosis,easily recrudescence and large mortality. Most patients are resistant to chemotherapyand radiotherapy at present. With increasing improvement of researching tumormethods, the molecular mechanisms of cancer are investigated clearly. It is hoped thatutilization of the RNAi method will help to inhibit target gene expression and inducetumor apoptosis. However, it is a serious challenge to researchers how to deliver geneinto tumor tissues safely and effectively. In recent years, nanotechnology has provideda novel approach to researching gene transmission and also new hope in treatingcancer. Thus, it is important to develop better gene delivery system for treatmentcancer.
Graphene, a novel and promising nanomaterial, has attracted tremendousattention due to its remarkable electronic, mechanical, and thermal properties.Functional graphene oxide （GO）was often applied in biomedical field. More andmore researchers had demonstrated that modified graphene oxide could improve genetransfection efficiency and biocompatibility in physiological environments.Polythyleneimine（PEI）was widely used for gene delivery, due to binding DNAability, protection of DNA against enzymatic degradation and high endosomal escape.GO-PEI functionalized with polyethylene glycol (PEG) had exhibited high solubilityand stability in physiological solutions. In this work, we applied GO-PEI-PEG asStat3siRNA gene delivery.
Signal transducer and activator of transcription3（Stat3）has a crucial role insignal transduction. Stat3is closely related to cell proliferation, differentiation andapoptosis. Constitutive activation of Stat3is the most intimately linked to tumourgenesis. Stat3has been found in an activated state in a wide variety of solidtumors, including malignant melanoma, liver cancer, and prostate cancer. Aberrantlyactive Stat3promotes proliferation, inhibits apoptosis, and influences cell cycleprogression. In previous work, we used a DNA vector-based Stat3-specific RNAiapproach to block Stat3signaling, and successfully inhibited of Stat3overexpressionin liver cancer and prostate cancer. But RNAi approach can not completely blocktarget gene expression in mammalian cells. Photothermal therapy has advantage overchemotherapy and radiotherapy, which has no toxicity or inhibits tumor growthinduction of cell apoptosis. Thus, we show much better therapeutic effficacy usingtargeted delivery in cancer treatment. Photothermal therapy and gene therapy havecombined effect on induction of tumor cell apoptosis.
We apply GO-PEI-PEG as a nanovehicle for Stat3-specific RNAi gene delivery tobetter kill tumor cells. We then utilize the strong optical absorbance of GO in theinfrared region for in vivo photothermal therapy, achieving ultraefficient tumorablation after intratumor administration of GO. Furthermore, in this work, we explorethe antitumor effect and therapeutic mechanism of functional GO carriedStat3-specific RNAi in malignant melanoma mouse model.
Objective: To evaluate the antitumor effect on Stat3-specific RNAi delivered byGO-PEI-PEG in mouse malignant melanoma and explore the antitumor underlyingmechanism in vitro and in vivo.
Method: We chemically synthetize GO-PEI-PEG nanovehicle. Nansheet sizes andzeta potential were determined by employing dynamic light scattering methodology.The morphology and size of nanosheets were imaged tapping-mode atomic forcemicroscopy (AFM). To determine stability of GO and GO-PEI-PEG nanosheets, thetwo samples were separately put into PBS, DMEM medium and serum at roomtemperature. To study Stat3-specific siRNA plasmid loading onto GO-PEI–PEGcomplexes, a gel electrophoresis assay was performed after incubation ofGO-PEI-PEG with si-Stat3plasmid at different mass ratios. The cytotoxicity ofGO-PEI-PEG to B16cells was evaluated by a CCK8assay. B16cells were transfectedwith si-Stat3plasmid delivered by GO-PEI-PEG. Tumor cell cycle was detected by PIassay. RT-PCR and Western blot analysis to detect the expression of Stat3relatedgenes and proteins changes were performed. To study the effects of si-Stat3on malignant melanoma growth in vivo, we developed a C57BL/6mouse tumorxenograft model. The morphology of apoptosis cells and GO-PEI-PEG was observedby transmission electron microscopy. Immunohistochemical analyses for CD34andPCNA protein expressions were carried out. TUNEL analyses were carried out toobserve cell apoptosis of tumor. BUN, ALT and AST in the serum were measured byauto-chemistry analyzer. The morphology of organs was examined in every group byhematoxylin and eosin (H&E) method.
Result: The nanosheets of GO-PEI-PEG were successfully synthesized. The zetapotential and effective diameter of the GO-PEI-PEG were determined to be＋21.02mV and298.9nm, respectively. After binding si-Stat3plasmid, the zeta potential andeffective diameter of complexes were determined to be＋7.52mV and339.2nm.Significant stability of GO-PEI-PEG was observed in physiological solutions afterextensive incubation.
GO-PEI-PEG complexes are a safe and effective nanocarrier for delivery ofplasmid Stat3siRNA in vitro. Transfection with GO-PEI-PEG/si-Stat3-GFP to B16cells were visualized green fluorescent protein (GFP). The Stat3mRNA, proteinexpression and Stat3related protein decreased significantly in the transfected withGO-si-Stat3by RT-PCR and Western blot. Cell cycle was blocked S cycle of tumor.
The results in vivo indicated significant regression in tumor growth and tumorweight after plasmid-based Stat3siRNA delivered by GO-PEI-PEG with simultaneousinfrared irradiation on the tumor. The Stat3mRNA and protein expression decreasedsignificantly in the mice transfected with GO-si-Stat3. Moreover, Stat3related proteinexpressions had significant alteration. Apoptosis of tumor cells in the GO-si-Stat3group were observed though H&E staining and transmission electron microscopy.Proliferating cells and tumor microvessels decreased in the GO-si-Stat3group viaimmunohistochemistry assay. The BUN, AST and ALT levels in the blood were alsonormal. No noticeable toxic effect on organs was found and no GO-PEI-PEG wasdistributed in heart, liver, spleen, lung and kidney in each group.
Conclusion: GO-PEI-PEG is an excellent nanocarrier for effective delivery ofplasmid Stat3siRNA. We then utilize the strong optical absorbance of GO in theinfrared region for in vivo photothermal therapy, achieving ultraefficient tumorablation after intratumor administration of GO. Plasmid Stat3siRNA delivered by GO-PEI-PEG with simultaneous infrared irradiation on mouse malignant melanomasuppressed tumor growth.
近几年，一种极具应用前景和优异性能的新型纳米材料—石墨烯，以其独特的力学、电学、热学等理化性质引起了研究者极大的兴趣。功能化氧化石墨烯（Graphene Oxide, GO）被生物医学领域的研究者广泛应用。迄今，随着纳米技术的高速发展，越来越多的实验证明，经过表面修饰的GO可以提高基因的转染效率，在生理环境下稳定性好。聚乙烯亚胺（polythyleneimine, PEI）是目前应用较多的阳离子基因传递载体，在生理环境下表面带正电荷，与基因的结合能力强，保护基因免受核酸酶的降解，具有从内涵体逃逸的能力。聚乙二醇（PolyethyleneGlycol，PEG）修饰的氧化石墨烯可以提高GO-PEI稳定性和溶解性，延长复合物在体内的循环时间。所以本实验将采用GO-PEI-PEG反应形成的共聚物作为Stat3siRNA质粒的运载体。
在信号转导通路中，信号转导子和转录激活子3（Signal transducer andactivator of transcription，Stat3）与肿瘤的发生发展密切相关。研究表明，在前列腺癌、肝癌及恶性黑色素瘤等多种肿瘤，Stat3呈现过度表达。Stat3的异常表达通过促进肿瘤新生血管的形成、抑制肿瘤细胞凋亡、促进肿瘤细胞增殖，参与肿瘤的发生过程。因此阻断肿瘤细胞中Stat3通路，可以达到治疗肿瘤的目的。在实验室前期工作中，以RNA干涉技术原理为基础，成功构建Stat3-特异siRNA表达质粒，抑制了前列腺癌、肝癌Stat3基因的过度表达[1,2]，促进肿瘤细胞凋亡。但是，RNAi并不能完全阻断目的基因的表达[3]。光热疗在物理学和生物学具有独特的的优势，与放疗、化疗相比，无毒副作用，具有杀伤肿瘤细胞的作用，可以作为治疗肿瘤的一种辅助手段。因此，光热疗和基因治疗联合策略治疗肿瘤迫在眉睫。
本课题将氧化石墨烯、聚乙烯亚胺、聚乙二醇反应形成共聚物作为肿瘤基因Stat3-特异siRNA质粒的运载体，更好的抑制或杀死肿瘤细胞。同时利用氧化石墨烯吸收红外线的物理学性质，在红外线光照下引起肿瘤组织局部温度升高，从而杀伤肿瘤细胞，进一步探讨功能化氧化石墨烯携带Stat3-特异siRNA质粒对小鼠恶性黑色素瘤的治疗效果及作用机制。
目的：功能化氧化石墨烯携带Stat3-特异siRNA表达质粒体内、体外抑制小鼠恶性黑色素瘤生长及作用机制的探讨。
方法：采用化学法合成氧化石墨烯—聚乙烯亚胺—聚乙二醇（GO-PEI-PEG）基因载体。通过原子力学显微镜、动态光散射等方法，对GO-PEI-PEG粒径、形貌、表面电势等进行表征；研究GO-PEI-PEG在PBS溶液、血清和培养液中的分散性和稳定性。利用琼脂糖凝胶电泳方法，检测在不同质量比条件下，GO-PEI-PEG与si-Stat3结合能力。CCK8法检测GO-PEI-PEG对B16细胞毒性。流式细胞术评价GO-PEI-PEG运载si-Stat3质粒的转染效率。PI染色法检测肿瘤的细胞周期。RT-PCR和Western blot检测Stat3基因及蛋白表达水平；同时检测Stat3相关下游基因的表达情况。建立C57BL/6小鼠恶性黑色素瘤皮下移植瘤模型。透射电子显微镜观察凋亡细胞以及石墨烯的形貌；免疫组织化学方法检测CD34和PCNA的表达；TUNEL试剂盒检测肿瘤细胞凋亡情况；全自动生化分析仪检测各组小鼠血液中BUN、ALT和AST的水平；HE染色观察各组脏器的形态。
结果：成功合成GO-PEI-PEG纳米材料，其表面电势为＋21.02mV，平均粒径为298.9nm，GO-PEI-PEG与si-Stat3质粒结合后形成的复合物，表面电势下降到＋7.52mV，平均粒径增大至339.2nm，在PBS溶液、血清和培养液中具有良好的分散性和稳定性。
体外实验证明，GO-PEI-PEG是一种安全、有效的基因运载体，GO-PEI-PEG携带si-Stat3质粒转染B16细胞，能够观察到绿色荧光蛋白（GFP）的表达。Westernblot和RT-PCR结果显示，GO-PEI-PEG转染si-Stat3组细胞中Stat3基因被干涉，Stat3下游基因表达受到抑制。细胞周期结果显示，GO-PEI-PEG转染si-Stat3组肿瘤细胞周期阻滞在S期，阻止细胞周期的进程。
体内实验证明，GO-PEI-PEG携带Stat3-siRNA质粒联合光照（简称GO-si-Stat3组）显著抑制小鼠恶性黑色素瘤的生长。GO-si-Stat3组的肿瘤瘤重和体积低于其他各组，具有统计学差异。在基因水平和蛋白水平，检测到GO-si-Stat3组Stat3表达受到抑制，Stat3下游调控基因和相关蛋白表达减弱。透射电镜和HE染色观察到凋亡细胞。免疫组织化学结果表明，GO-si-Stat3组增殖的肿瘤细胞减少，微血管生成减少。各组小鼠血液中的BUN、ALT、AST浓度均在正常范围内。HE染色未发现异常的心、肝、脾、肺、肾形态。
结论：功能化氧化石墨烯是一种安全、有效的基因运载体，能够携带Stat3-特异siRNA表达质粒，显著抑制小鼠恶性黑色素瘤的生长。氧化石墨烯具有吸收红外线的物理学性质，联合红外线光照，与Stat3-特异siRNA表达质粒共同发挥抑制小鼠恶性黑色素瘤生长的作用。
Background: Malignant Melanoma is a common cancer, which has high aggressivemalignancy. Patients are effectively treated by surgery at early stages. However, theincidence rates of malignant melanoma in many countries are increasing evidently inrecent years. There are some important biological characteristics in malignantmelanoma, such as high aggressive malignancy, early metastasis, poor prognosis,easily recrudescence and large mortality. Most patients are resistant to chemotherapyand radiotherapy at present. With increasing improvement of researching tumormethods, the molecular mechanisms of cancer are investigated clearly. It is hoped thatutilization of the RNAi method will help to inhibit target gene expression and inducetumor apoptosis. However, it is a serious challenge to researchers how to deliver geneinto tumor tissues safely and effectively. In recent years, nanotechnology has provideda novel approach to researching gene transmission and also new hope in treatingcancer. Thus, it is important to develop better gene delivery system for treatmentcancer.
Graphene, a novel and promising nanomaterial, has attracted tremendousattention due to its remarkable electronic, mechanical, and thermal properties.Functional graphene oxide （GO）was often applied in biomedical field. More andmore researchers had demonstrated that modified graphene oxide could improve genetransfection efficiency and biocompatibility in physiological environments.Polythyleneimine（PEI）was widely used for gene delivery, due to binding DNAability, protection of DNA against enzymatic degradation and high endosomal escape.GO-PEI functionalized with polyethylene glycol (PEG) had exhibited high solubilityand stability in physiological solutions. In this work, we applied GO-PEI-PEG asStat3siRNA gene delivery.
Signal transducer and activator of transcription3（Stat3）has a crucial role insignal transduction. Stat3is closely related to cell proliferation, differentiation andapoptosis. Constitutive activation of Stat3is the most intimately linked to tumourgenesis. Stat3has been found in an activated state in a wide variety of solidtumors, including malignant melanoma, liver cancer, and prostate cancer. Aberrantlyactive Stat3promotes proliferation, inhibits apoptosis, and influences cell cycleprogression. In previous work, we used a DNA vector-based Stat3-specific RNAiapproach to block Stat3signaling, and successfully inhibited of Stat3overexpressionin liver cancer and prostate cancer. But RNAi approach can not completely blocktarget gene expression in mammalian cells. Photothermal therapy has advantage overchemotherapy and radiotherapy, which has no toxicity or inhibits tumor growthinduction of cell apoptosis. Thus, we show much better therapeutic effficacy usingtargeted delivery in cancer treatment. Photothermal therapy and gene therapy havecombined effect on induction of tumor cell apoptosis.
We apply GO-PEI-PEG as a nanovehicle for Stat3-specific RNAi gene delivery tobetter kill tumor cells. We then utilize the strong optical absorbance of GO in theinfrared region for in vivo photothermal therapy, achieving ultraefficient tumorablation after intratumor administration of GO. Furthermore, in this work, we explorethe antitumor effect and therapeutic mechanism of functional GO carriedStat3-specific RNAi in malignant melanoma mouse model.
Objective: To evaluate the antitumor effect on Stat3-specific RNAi delivered byGO-PEI-PEG in mouse malignant melanoma and explore the antitumor underlyingmechanism in vitro and in vivo.
Method: We chemically synthetize GO-PEI-PEG nanovehicle. Nansheet sizes andzeta potential were determined by employing dynamic light scattering methodology.The morphology and size of nanosheets were imaged tapping-mode atomic forcemicroscopy (AFM). To determine stability of GO and GO-PEI-PEG nanosheets, thetwo samples were separately put into PBS, DMEM medium and serum at roomtemperature. To study Stat3-specific siRNA plasmid loading onto GO-PEI–PEGcomplexes, a gel electrophoresis assay was performed after incubation ofGO-PEI-PEG with si-Stat3plasmid at different mass ratios. The cytotoxicity ofGO-PEI-PEG to B16cells was evaluated by a CCK8assay. B16cells were transfectedwith si-Stat3plasmid delivered by GO-PEI-PEG. Tumor cell cycle was detected by PIassay. RT-PCR and Western blot analysis to detect the expression of Stat3relatedgenes and proteins changes were performed. To study the effects of si-Stat3on malignant melanoma growth in vivo, we developed a C57BL/6mouse tumorxenograft model. The morphology of apoptosis cells and GO-PEI-PEG was observedby transmission electron microscopy. Immunohistochemical analyses for CD34andPCNA protein expressions were carried out. TUNEL analyses were carried out toobserve cell apoptosis of tumor. BUN, ALT and AST in the serum were measured byauto-chemistry analyzer. The morphology of organs was examined in every group byhematoxylin and eosin (H&E) method.
Result: The nanosheets of GO-PEI-PEG were successfully synthesized. The zetapotential and effective diameter of the GO-PEI-PEG were determined to be＋21.02mV and298.9nm, respectively. After binding si-Stat3plasmid, the zeta potential andeffective diameter of complexes were determined to be＋7.52mV and339.2nm.Significant stability of GO-PEI-PEG was observed in physiological solutions afterextensive incubation.
GO-PEI-PEG complexes are a safe and effective nanocarrier for delivery ofplasmid Stat3siRNA in vitro. Transfection with GO-PEI-PEG/si-Stat3-GFP to B16cells were visualized green fluorescent protein (GFP). The Stat3mRNA, proteinexpression and Stat3related protein decreased significantly in the transfected withGO-si-Stat3by RT-PCR and Western blot. Cell cycle was blocked S cycle of tumor.
The results in vivo indicated significant regression in tumor growth and tumorweight after plasmid-based Stat3siRNA delivered by GO-PEI-PEG with simultaneousinfrared irradiation on the tumor. The Stat3mRNA and protein expression decreasedsignificantly in the mice transfected with GO-si-Stat3. Moreover, Stat3related proteinexpressions had significant alteration. Apoptosis of tumor cells in the GO-si-Stat3group were observed though H&E staining and transmission electron microscopy.Proliferating cells and tumor microvessels decreased in the GO-si-Stat3group viaimmunohistochemistry assay. The BUN, AST and ALT levels in the blood were alsonormal. No noticeable toxic effect on organs was found and no GO-PEI-PEG wasdistributed in heart, liver, spleen, lung and kidney in each group.
Conclusion: GO-PEI-PEG is an excellent nanocarrier for effective delivery ofplasmid Stat3siRNA. We then utilize the strong optical absorbance of GO in theinfrared region for in vivo photothermal therapy, achieving ultraefficient tumorablation after intratumor administration of GO. Plasmid Stat3siRNA delivered by GO-PEI-PEG with simultaneous infrared irradiation on mouse malignant melanomasuppressed tumor growth.
引文
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