靶向癌胚抗原载药纳米超声造影剂体外抑制卵巢癌细胞生长

摘要

Objective To prepare carcino-embryonic antigen (CEA) targeted and paclitaxel loaded phase-shifting PLGA nanoparticles (Ab-PTX-NPs),and investigate the targeting capability and inhibition to the ovarian cancer cell in vitro.Methods Single-emulsion/solvent evaporation (O/W) and carbodiimide method were used to prepare the Ab-PTX-NPs.The size of nanoparticles was determined by Malvern analyzer.The encapsulation and drug loaded efficiency of paclitaxel were detected by high performance liquid chromatography.And the drug release characteristics was measured by dialysis method in constant temperature shaker.The targeting ability of Ab-PTX-NPs to the ovarian cancer SKOV3 cell was evaluated by the laser scanning confocal microscope and flow cytometry.And the inhibition ability of Ab-NPs was investigated by the CCK-8 assays.Results The size of Ab-PTX-NPs was (397.70±99.95)nm.The encapsulation efficiency and drug loading capacity of PTX were (67.26±4.15) ％ and (6.31±0.39) ％,respectively.The conjugating rate of Anti-CEA antibody was (92.74 ± 5.75) ％.The targeting study in vitro showed that such a number of contrast agents landed around the SKOV3 cells in targeting group,and the mean fluorescence intensity of ovarian cells in targeting group was significantly higher than other groups (P＜0.05).After 24 h,the viability rate of ovarian cells in targeting group was lower than the non-target group (P＜0.05),only higher than that of the pure PTX group (P＜0.05).But there was no significant difference between the targeting group and the pure PTX group (P＞0.05) at 48 h.Conclusion The CEA targeted and paclitaxel loaded phase-shifting PLGA nanoparticles are successfully prepared.It can enhance ultrasound imaging well after activated by LIFU.With high drug-loading efficiency and fast drug release velocity,the Ab-PTX-NPs appeares great targeted ability.%目的 制备靶向癌胚抗原(CEA)载药相变型PLGA纳米粒(Ab-PTX-NPs),探讨该纳米粒的体外寻靶及抑制肿瘤细胞生长的效能.方法 乳化溶剂挥发法联合碳二亚胺法制备靶向CEA载紫杉醇纳米粒(Ab-PTX-NPs),采用马尔文粒径分析仪检测其粒径大小.采用高效液相色谱法检测紫杉醇包封率及载药量,恒温摇床透析法检测该纳米粒体外释药特征.激光共聚焦显微镜及流式细胞术观察该纳米粒体外寻靶情况,CCK-8试剂检测卵巢癌细胞存活率.结果 制备的Ab-PTX-NPs粒径为(397.70±99.95) nm,紫杉醇包封率及载药量分别为(67.26±4.15)％和(6.31±0.39)％.抗CEA单克隆抗体与纳米粒连接率为(92.74±5.75)％.共聚焦显微镜下观察到靶向组卵巢癌SKOV3细胞周围见较多造影剂黏附,流式细胞术测得靶向组细胞平均荧光强度明显高于其余各组(P＜0.05),CCK-8试剂法测得靶向组在24 h时,细胞存活率高于纯药组(P＜0.05),而低于无靶组(P＜0.05),当48 h时,靶向组与纯药组细胞存活率差异无统计学意义(P＞0.05).结论 成功制备靶向CEA载药相变型PLGA纳米粒,该纳米粒经低功率聚焦超声治疗仪(LIFU)致相变后可明显增强超声显影,且载药量高,释药快,靶向能力强.