整合素受体和细胞穿膜肽共修饰紫杉醇脂质体抑制食管癌Ec9706细胞的研究

The Inhibition Effect of RGD and R8 Co-modified Paclitaxel Loaded Liposome on the Esophagus Carcinoma EC9706 Cells

目的:制备整合素受体RGD和细胞穿膜肽R8共修饰载紫杉醇(PTX)脂质体(RGD/R8-LP-PTX),对其理化性质进行表征,并观察脂质体与食管癌Ec9706细胞的亲和力和增殖抑制作用。方法:采用薄膜分散法制备RGD/R8-LP-PTX,观察脂质体的粒径,电位以及包封率;通过定量细胞摄取实验观察食管癌Ec9706细胞对RGD/R8-LP的摄取效率以及对脂质体摄取的影响因素。定性共聚焦实验观察肿瘤细胞对脂质体的摄取。MTT实验观察RGD/R8-LP-PTX对食管癌Ec9706细胞的细胞毒性;构建食管癌Ec9706细胞肿瘤球模型,观察脂质体对肿瘤球的生长抑制能力。结果:RGD/R8-LP-PTX的粒径在124.8±9.4 nm,电位为21.35±3.55 m V。食管癌Ec9706细胞对RGD/R8-LP的摄取以及RGD/R8-LP-PTX对食管癌Ec9706细胞的增殖抑制率具有时间依赖性;食管癌Ec9706细胞对RGD/R8-LP的摄取效率显著高于R8-LP、RGD-LP和LP,差异有统计学意义(P<0.01);在给药48 h后,R8-LP-PTX、RGD-LP-PTX和LP-PTX的细胞存活率分别是RGD/R8-LP-PTX组的1.6倍、1.7倍和2.2倍,差异有统计学意义(P<0.01)。给药7天后,生理盐水组肿瘤球持续生长,体积增大1.48倍,LP-PTX组肿瘤球体积增大到原体积的1.12倍,RGD/R8-LP-PTX组、R8-LP-PTX组和RGD-LP-PTX组肿瘤球体积减小到原体积的36%、59%和64%,差异具有统计学意义(P<0.01)。结论:整合素受体RGD和细胞穿膜肽R8共修饰载紫杉醇(PTX)脂质体能够有效穿透肿瘤细胞膜进入肿瘤细胞,是一种有效的食管癌化疗靶向给药系统。

Objective： To prepare RGD and R8 co-modified paclitaxel loaded liposome（RGD/R8-LP-PTX）for EC9706 cells’ treatment. Methods： The co-modified liposome was prepared by film-ultrasonic method. The appearance, particle size,Zeta potential were evaluated. The cellular uptake by EC9706 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/R8-LP-PTX was evaluated by MTT assay. Tumor spheroids were used to evaluate anti-tumor ability of RGD/R8-LP-PTX in vitro. Results：The particle diameter of the co-modified liposome was 124. 8 ± 9. 4 nm with the Zeta potential of 21. 35 ± 3. 55mV. The uptake of the RGD/R8-LP by Ec9706 cells and the prolif-eration inhibition rate of RGD/R8-LP-PTX to Ec9706 cells were both found in a time-dependent manner. The result demon-strated that the co-modified liposome uptaken by EC9706 were 2. 1, 2. 6 times higher than that of R8-LP and RGD-LP, re-spectively. The MTT assay demonstrated the cell viability of R8-LP-PTX,RGD-LP-PTX and LP-PTX were 1. 6, 1. 7 and 2. 2 times higher than that of RGD/R8-LP-PTX respectively. Tumor spheroids continued to grow in size and volume in saline 148% of the primary volume while the tumor spheroid volumes changed into nearly 36%, 59% and 64% of the original volumes when treated with R8-LP-PTX,RGD-LP-PTX and LP-PTX respectively（P〈0. 01）. Conclusion： The RGD/R8-LP-PTX can penetrate the tumor cell membrane effectively ,it might serve as a promising delivery system of antitumor drugs against esophagus carcinoma.