叶酸靶向载顺铂羧甲基-β-环糊精纳米复合物的制备及对鼻咽癌的体外抑制效应  

The Preparation of Folate-targeted Carboxymethyl-β-cyclodextrin Nanocomposites Loaded with Cisplatin and Its Evaluation of Inhibitory Effect on Nasopharyngeal Carcinoma in vitro

作　　者：刘涛[1] 马栋[2] 柯波[3] 谢民强[1] 

机构地区：[1]南方医科大学珠江医院耳鼻咽喉头颈外科,广州510282 [2]暨南大学生物医学工程系,广州510630 [3]江西省人民医院江西血液肿瘤细胞生物学重点实验室,南昌330006

出　　处：《中国细胞生物学学报》2014年第1期60-68,共9页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81372477;81260406);教育部高等学校博士学科点专项科研基金(批准号:20114433110001);江西省自然科学基金(批准号:20122BAB205068)资助的课题~~

摘　　要：为达到鼻咽癌(nasopharyngeal carcinoma,NPC)的靶向化疗,该研究通过酰胺化反应和配位偶联技术制备叶酸(folic acid,FA)分子靶向载顺铂(cisplatin,CDDP)羧甲基-β-环糊精(carboxymethyl-β-cyclodextrin,CM-β-CD)纳米复合物(FA-CM-β-CD-CDDP),采用邻苯二胺(o-phenylenediamine,OPDA)比色法检测复合物中CDDP含量,紫外分光光谱检测FA含量,透射电镜观察复合物形态,激光粒度仪测定复合物粒径大小。荧光显微镜观察NPC叶酸受体(folate receptor,FR)阳性HNE-1细胞及FR阴性CNE-2细胞对偶联FITC的复合物的吞噬及OPDA比色法检测细胞内CDDP的浓度。通过MTT法、集落形成实验和流式细胞术检测复合物对HNE-1细胞增殖能力和凋亡的影响。研究结果显示,复合物中偶联的FA和CDDP浓度分别为340μg/mL和2 mg/mL,CDDP包封率达20.00%,复合物粒径均匀且大小为157.8 nm。HNE-1细胞内见较多FITC,细胞内CDDP浓度为6.24 ng/mL,而CNE-2细胞内FITC较少,细胞内CDDP浓度仅约2.01 ng/mL。HNE-1生长抑制率在24 h明显高于对照组(CM-β-CD-CDDP),其IC50(4.80μg/mL)明显低于对照组(6.97μg/mL),但当所载的CDDP终浓度达到16.00μg/mL时,两组抑制率均达到80%以上;作用48 h两组抑制率无明显差异。在24 h,当复合物的CDDP终浓度为1.00μg/mL时,HNE-1的集落形成率为33.21%,明显低于对照组(52.27%)。当复合物的CDDP终浓度为0.25μg/mL和1.00μg/mL时,HNE-1的凋亡率分别达12.65%和22.35%,明显高于对照组(6.91%和14.21%)。研究结果表明,成功构建的FA-CM-β-CD-CDDP纳米复合物能够靶向抑制FR阳性的NPC细胞增殖并促进其凋亡。In order to achieve was prepared by amidation reaction and targeted chemotherapy for NPC, FA-CM-β-CD-CDDP nanocomposites ligand coupling technology in this research. The concentration of CDDP, concentration of FA, morphology and particle size of nanocomposites were detected by OPDA colourimetry, UV spectroscopy, transmission electron microscopy and laser particle detector, respectively. The phagocytic effects of NPC FR expressing positive HNE-1 cells and FR expressing negative CNE-2 cells on FA-CM-β-CD-CDDP coupling with FITC were observed using fluorescence microscope and the concentration of CDDP in cells was detected by OPDA colourimetry. The effects of nanocomposites on HNE-1 cells proliferation and apoptosis were measured with MTT, colony forming experiment and flow cytometry. Results showed that concentration of FA and CDDP coupled with nanocomposites were 340 μg/mL and 2 mg/mL, respectively, and the encapsulation efficiency of CDDP was 20.00%. FA-CM-β-CD-CDDP nanocomposites was in uniform dispersion, and its average particle size was 157.8 nm. More FITC was seen in HNE-1 cells, concentration of CDDP in cells was 6.24 ng/mL, but less FITC was seen in CNE-2 cells, and concentration of CDDP in cells was only 2.01 ng/mL. The growth inhibiting ratio of HNE-1 cells in FA-CM-β-CD-CDDP group was significantly increased compared to control group （CM-β-CD- CDDP） after cells were treated for 24 h. IC50 （4.80 μg/mL） of CDDP in FA-CM-β-CD-CDDP group was significantly decreased compared to control group （6.97 μg/mL）. Cell growth inhibition rate of the two groups all were above 80% when the final concentration of CDDP of the two kinds of nanocomposites reached 16.00μg/mL, and cell growth inhibition rate of the two groups had no significant difference after 48 h. Colony forming rate （33.21%） of HNE-1 cells in FA-CM-β-CD-CDDP group was significantly lower than that of control group （52.27%） when the final concentration of CDDP in two groups were 1.00 μg/mL, and apoptosis rates of liNE-1 cell

关 键 词：叶酸 分子靶向 纳米复合物 鼻咽癌 顺铂 

分 类 号：R944[医药卫生—药剂学] R96[医药卫生—药学]