石墨烯纳米载药体系的制备及对肿瘤细胞的杀伤作用

doi:10.19894/j.issn.1000-0518.200302

摘要/Abstract

摘要： 通过聚乙二醇(PEG)及分支型聚乙烯亚胺(bPEI)对纳米氧化石墨烯(NGO)修饰作为大分子载药基底的载药平台,增加了NGO的水溶性及其对蛋白的吸附作用,随后分别负载抗癌药物顺铂(CDDP)和低温乙醇法分离提纯的肿瘤患者血清形成能够特异性富集于鼻咽癌细胞的大分子石墨烯纳米载药体系。通过紫外-可见光谱和傅里叶变换红外光谱表征结果证实NGO-PEG-bPEI-CDDP载药体系制备成功,NGO-PEG-bPEI对CDDP的载药率为34.6%。聚丙烯酰胺凝胶电泳(SDS-PAGE)表明对NGO-PEG-bPEI-CDDP-Antibody大分子纳米载药体系中特异性抗体蛋白的强烈吸附作用,该纳米复合物能够特异性地富集在肿瘤细胞部位,对人鼻咽癌细胞(CNE-1)细胞系的识别极其敏感。细胞毒性实验(MTT)检测实验结果表明:在相同剂量(质量浓度)、相同作用时间的情况下,NGO-PEG-bPEI-CDDP-Antibody大分子纳米石墨烯载药体系兼具对人口腔鳞癌细胞(KB)、CNE-1杀伤作用和避免正常细胞的额外损伤。 NGO-PEG-bPEI-CDDP-Antibody纳米载药体系不仅能够通过特异性识别将药物富集于病灶区降低正常细胞损害,还能够有效杀伤癌细胞,降低化疗药物使用剂量,是一种很有前景的大分子纳米载药体系。

关键词: 纳米氧化石墨烯, 大分子载药体系, 肿瘤特异性抗体, 肿瘤富集作用

Abstract: In this study, a nano-graphene oxide (NGO) was modified by polyethylene glycol (PEG) and branched polyethyleneimine (bPEI) as a drug loading platform of macromolecular drugs, which increases the water solubility of NGO and the adsorption of proteins. Then, tumor patient serum was prepared by loading anti-cancer drug cis-diamminedichloroplatinum (CDDP) and low-temperature ethanol, respectively, to form the graphene nanoparticle drug delivery system. The ultraviolet-visible and Fourier transform infraredspectroscopy characterization results demonstrate that the NGO-PEG-bPEI-CDDP drug loading system is successfully prepared, and the drug loading rate of NGO-PEG-bPEI to CDDP is 34.6%. Polyacrylamide gel electrophoresis (SDS-PAGE) demonstrates strong adsorption of specific antibody proteins in the NGO-PEG-bPEI-CDDP-Antibody macromolecular nano-loading system, and the nanocomposite can be specifically enriched in tumors. Cell sites are extremely sensitive to the recognition of CNE-1 cell lines. Cytotoxicity test (MTT) results demonstrate that the drug delivery system of NGO-PEG-bPEI-CDDP-Antibody macromolecular nano graphene can kill KB and CNE-1 cells and avoid additional damage to normal cells at the same concentration and time. The NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system can not only enrich the drug in the lesion area through specific recognition, reduce normal cell damage, but also effectively kill cancer cells and reduce the dose of chemotherapy drugs, which is a very promising macromolecular nano-drug loading system.

Key words: Nano-graphene oxide, Macromolecular drug loading system, Tumor-specific antibody, Tumor enrichment

中图分类号:

O611

张雅静, 王晓辉, 徐亚娟, 刘文书, 赵丽辉. 石墨烯纳米载药体系的制备及对肿瘤细胞的杀伤作用[J]. 应用化学, 2021, 38(6): 693-703.

ZHANG Ya-Jing, WANG Xiao-Hui, XU Ya-Juan, LIU Wen-Shu, ZHAO Li-Hui. Preparation of Graphene Nano-Drug Carrier System and Its Killing Effect on Tumor Cells[J]. Chinese Journal of Applied Chemistry, 2021, 38(6): 693-703.

参考文献

[1] LIN Z R, XU Y Y, ZHANG T. A theoretical exploration into durability of carbon nanotubes cement-based materials[J]. Low Temp Architect Technol, 2011, 2(3): 996-1000.
[2] BANERJEE A N. Graphene and its derivatives as biomedical materials: future prospects and challenges[J]. Interface Focus, 2018, 8(3): 1-22.
[3] RADUNOVIC M, DE C M, DE M P, et al. Graphene oxide enrichment of collagen membranes improves DPSCs differentiation and controls inflammation occurrence[J] . J Biomed Mater Res Part A, 2017, 105(8): 2312-2320.
[4] LI Y, LU Q, LIU H, et al. Antibody-modified reduced graphene oxide films with extreme sensitivity to circulating tumor cells[J]. Adv Mater, 2015, 27(43): 6848-6854.
[5] ZHANG C, LIU Z, ZHENG Y, et al. Glycyrrhetinic acid functionalized graphene oxide for mitochondria targeting and cancer treatment in vivo[J]. Small, 2018, 14(4): 1703306.
[6] 梁爽, 韩雷强, 张娜. 结构修饰的基于聚乙烯亚胺的载体在基因递送方面的研究进展[J]. 药物生物技术, 2017, 10(4): 55-61.
LIANG S, HAN L Q, ZHANG N. Research progress of structurally modified polyethyleneimine-based carriers in gene delivery[J]. Pharm Biotechnol, 2017, 10(4): 55-61.
[7] 武叙, 李晓宇, 姚翀, 等. BPEI/miRNA, PAMAM/miRNA和PEI/miRNA治疗前列腺癌作用研究[J]. 中国药学杂志, 2015, 50(9): 768-774.
WU X, LI X Y, YAO C, et al. Effect of BPEI/miRNA, PAMAM/miRNA and PEI/miRNA delivery system on the treatment of prostate cancer[J]. J Chinese Pharm, 2015, 50(9): 768-774.
[8] OU M, WANG XL, XU R, et al. Novel biodegradable poly(disulfide amine)s for gene delivery with high efficiency and low cytotoxicity[J]. Bioconjugate Chem, 2008, 19(3): 626-633.
[9] 杨传孝, 余梦雯, 宋朵朵, 等. 甲醛功能化聚乙烯亚胺-罗丹明B酰肼比率及可视化荧光测定Cr(Ⅵ)[J]. 高等学校化学学报, 2016, 37(5): 852-859.
YANG C X, YU M W, SONG D D, et al. Modification of formaldehyde-functionalized polyethyleneimine-rhodamine B hydrazide and its visual fluorescence spectrometry for Cr(VI)[J]. Chem J Chinese Univ, 2016, 37(5): 852-859.
[10] ZHANG L, LU Z, ZHAO Q, et al. Enhanced chemotherapy efficacy by sequential delivery of siRNA and anticancer drugs using PEI-grafted graphene oxide[J]. Small, 2011, 7(4): 460-464.
[11] ZHOU T, ZHOU X, XING D. Controlled release of doxorubicin from graphene oxide based charge-reversal nanocarrier[J]. Biomaterials, 2014, 35 (13): 4185-4194.
[12] WENG Y, JIANG B, YANG K, et al. Polyethyleneimine-modified graphene oxide nanocomposites for effective protein functionalization[J]. Nanoscale, 2015, 7(34): 14284-14292.