转铁蛋白与细胞穿膜肽共修饰脂质体抑制视网膜母细胞瘤的作用研究

Transferrin and transcriptional activator protein co-modified docetaxel loaded liposome inhibiting retinoblastoma

目的采用薄膜分散法制备转铁蛋白(transferrin,TF)与细胞穿膜肽(transcriptional activator protein,TAT)共修饰载多西紫杉醇(docetaxel,DOC)脂质体(TF/TAT-LP-DOC),探讨其对视网膜母细胞瘤(HXO-RB44)的靶向治疗作用。方法采用薄膜分散法制备TF/TAT-LP-DOC,并对其进行表征。通过MTT实验考察脂质体对HXO-RB44细胞的毒性,流式细胞仪检测HXORB44细胞对不同脂质体的摄取效率。构建HXO-RB44细胞肿瘤球模型,研究TF/TAT-LP-DOC对实体肿瘤的生长抑制作用。结果所制备的TF/TATLP-DOC粒径为(115.8±8.5)nm,Zeta电位为(23.58±3.65)m V,DOC的包封率为85.8%。MTT检测结果显示,LP-DOC、TFLP-DOC、TATLP-DOC和TF/TATLP-DOC组HXO-RB44细胞存活率分别为66.5%、43.6%、39.4%和18.9%,差异有统计学意义(P<0.01)。与LP-DOC、TFLP-DOC和TATLP-DOC组比较,TF/TATLP-DOC组的肿瘤细胞存活率显著低于其他脂质体组,差异均有统计学意义(均为P<0.01)。TF/TATLP-DOC组细胞存活率随时间的延长而降低,差异有统计学意义(P<0.01)。HXO-RB44细胞对TF/TATLP的摄取效率分别是TFLP、TATLP和LP的2.65倍、2.32倍和3.86倍,差异均有统计学意义(均为P<0.01)。TFLP和TATLP的细胞摄取效率高于LP,差异均有统计学意义(均为P<0.01)。相同脂质体在4 h的摄取效率高于2 h,差异有统计学意义(P<0.01)。给药7 d后,生理盐水组肿瘤球持续生长,体积增大1.44倍,LPDOC组肿瘤球体积增大到原体积的1.14倍,TF/TAT-LP-DOC组、TATLP-DOC组和TFLP-DOC组肿瘤球体积减小到原体积的35%、62%和58%,与LP-DOC、TFLP-DOC和TATLP-DOC组比较,TF/TATLP-DO组肿瘤球体积显著小于其他脂质体组,差异均有统计学意义(均为P<0.01)。肿瘤球体积随着时间的延长而减小,差异有统计学意义(P<0.01)。结论 TF/TATLP-DOC制备工艺简单,与HXO-RB44细胞具有良好的亲和性,是一种潜在高效的肿瘤靶向给药系统。

Objective To prepare transferrin （TF）and transcriptional activator protein （TAT） co-modified docetaxel loaded liposome （ TF/TATLP-DOC ）, and discuss its targeting therapy for retinoblastoma. Methods The co-modified liposome was pre- pared by film-ultrasonic method. The particle size, Zeta potential were evaluated. The cellular uptake by HXO-RB44 cells in vitro was used to evaluate the targeting efficien- cy. The anti-proliferation efficiency of TF/TATLP- DOC was evaluated by MTY assay. Results The particle diameter of the co-modified liposome was （ 115. 8 ± 8.5 ） nm with the Zeta potential of （23.58 ± 3.65 ） mV. The entrapment efficiency of DOC was 85. 8%. The survival rate of LP-DOC, TFLP-DOC, TATLP-DOC and TF/TATLP-DOC for HXO-RB44 cells were 66.5% ,43.6% ,39.4% and 18.9% ,respectively（P 〈0.01 ）. Com- pared with LP-DOC, TFLP-DOC, TATLP-DOC group, the survival rate of HXO-RB44 cells in TF/TATLP-DOC group was lower （all P 〈 0.01 ） ,which decreased with the time pro- long （P 〈 0.01 ）. The co-modified liposome uptake for TF/TATLP by HXO-RB44 were 2.65,2.32 and 3.86 times higher than that of TFLP,TATLP and LP,respectively（all P 〈0.01 ） ,and the co-modified liposome uptake for TFLP and TATLP were higher than that for LP （ all P 〈 0.01 ）. The uptake for same liposome at 4 hours was high- er than that at 2 hours （P 〈0.01 ）. After 7 days,the inhibition rate of NS,LP-DOC,TFLP-DOC,TATLP-DOC and TF/TATLP- DOC for HXO-RB44 tumor spheroids size were 144% , 114% ,35% ,62% and 58% ,the TF/TATLP-DOC group was smaller obviously than the LP-DOC ,TFLP-DOC ,TATLP-DOC group（ all P 〈 0.01 ）. The tumor spheroids size was decreased with the time prolong （P 〈 0.01 ）. Conclusion The establishment for TF/TATLP-DOC is simple with a well compatibility with HXO-RB44 cells, which might serve as a promising cancer delivery system of antitumor drugs.