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背景:结合肿瘤的分子靶向技术和纳米技术,制备出一种新的具有靶向作用的纳米载体,以达到对肿瘤更好靶向作用的目的。目的:制备表皮生长因子偶联牛血清白蛋白纳米载体,联合核素标记的c-erbB2反义寡脱氧核苷酸,体外观察人乳腺癌SK-BR3细胞对其摄取情况。设计、时间及地点:对比观察实验,于2006-09/2008-03在重庆医科大学生物化学和分子生物学教研室,重庆医科大学分子医学与肿瘤研究中心,重庆医科大学放射医学教研室完成。材料:牛血清白蛋白(生物技术级)由美国Amresco公司提供,表皮生长因子由英国PeproTech EC LTD提供,125I由成都中核高通同位素股份有限公司提供,寡脱氧核苷酸由上海生工生物工程技术服务有限公司提供。方法:采用超声乳化-化学交联及羧和反应制备表皮生长因子偶联白蛋白靶向纳米载体,通过核素标记示踪技术检测表皮生长因子偶联白蛋白靶向纳米载体的相关性质。主要观察指标:测量表皮生长因子靶向纳米载体荷载c-erbB2反义寡脱氧核苷酸的载药量、包封率及释药率,人乳腺癌SK-BR3细胞对表皮生长因子靶向纳米载体的摄取率及滞留率。结果:表皮生长因子靶向纳米载体包载125I标记的反义寡脱氧核苷酸组的摄取率及滞留率高于正义寡脱氧核苷酸组和无义寡脱氧核苷酸组。同时c-erbB2寡脱氧核苷酸使用纳米载体荷载组的摄取率及滞留率均高于未使用纳米载体组,差异有显著性意义(P<0.05)。结论:125I标记的表皮生长因子靶向纳米载体能够提高乳腺癌SK-BR3细胞对c-erbB2反义寡脱氧核苷酸的摄取和滞留,能达到更好的靶向作用。
BACKGROUND: Combined with the molecular targeted technology of tumor and nanotechnology, a new targeting nanocarrier has been prepared to achieve the goal of better targeting of tumor. OBJECTIVE: To prepare epidermal growth factor-conjugated bovine serum albumin nanocarriers and combine with nuclide-labeled c-erbB2 antisense oligodeoxynucleotides to observe the uptake of human breast cancer SK-BR3 cells in vitro. DESIGN, TIME AND SETTING: The comparative observation experiment was performed at the Department of Biochemistry and Molecular Biology, Chongqing Medical University from 2006-09 / 2008-03, the Department of Molecular Medicine and Cancer Research, Chongqing Medical University, and the Department of Radiation Medicine, Chongqing Medical University. MATERIALS: Bovine serum albumin (biotechnology grade) was supplied by American Amresco Corporation. Epidermal growth factor was provided by PeproTech EC LTD. 125I was supplied by Chengdu Zhongkon Qualcomm Isotope Co., Ltd. The oligodeoxynucleotides were obtained from Shanghai Bioengineering Engineering Services Limited provide. METHODS: Epidermal growth factor coupled albumin-targeting nanocarriers were prepared by phacoemulsification-chemical crosslinking and carboxylation. The related properties of epidermal growth factor-conjugated albumin-targeting nanocarriers were detected by radionuclide labeling. MAIN OUTCOME MEASURES: The drug loading, entrapment efficiency and drug release rate of c-erbB2 antisense oligodeoxynucleotide loaded on epidermal growth factor-targeted nanocarriers were measured, and the effect of SK-BR3 cells on the proliferation of epidermal growth factor Carrier uptake rate and retention rate. Results: The uptake rate and retention rate of epidermal growth factor (EGF) targeting nanocarriers on 125I-labeled antisense oligodeoxynucleotides were higher than those of oligodeoxynucleotides and non-sense oligodeoxynucleotides. At the same time, the uptake rate and retention rate of c-erbB2 oligodeoxynucleotides in the nanocarrier-loaded group were significantly higher than those in the non-nanocarrier group (P <0.05). Conclusion: 125I-labeled epidermal growth factor-targeting nanocarriers can enhance the uptake and retention of c-erbB2 antisense oligodeoxynucleotides in breast cancer SK-BR3 cells and achieve better targeting.