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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 / R8-LP-PTX liposomes, which are integrin receptor RGD and cell penetrating peptide R8, to characterize their physico-chemical properties and observe the relationship between liposomes and esophageal cancer Ec9706 Cell affinity and proliferation inhibition. Methods: RGD / R8-LP-PTX was prepared by membrane dispersion method. The size, potential and entrapment efficiency of liposomes were observed. The uptake efficiency of RGD / R8-LP in esophageal carcinoma Ec9706 cells by quantitative cell uptake assay Influencing factors of liposome uptake. Qualitative confocal experiments were performed to observe the uptake of liposomes by tumor cells. The cytotoxicity of RGD / R8-LP-PTX on esophageal cancer cell line Ec9706 was observed by MTT assay. The esophageal cancer Ec9706 cell model was constructed and the inhibitory effect of liposome on tumor growth was observed. Results: The particle size of RGD / R8-LP-PTX was 124.8 ± 9.4 nm with a potential of 21.35 ± 3.55 mV. The uptake of RGD / R8-LP by esophageal cancer Ec9706 cells and the inhibition of the proliferation of esophageal cancer Ec9706 cells by RGD / R8-LP-PTX were time-dependent. The uptake efficiency of RGD / R8-LP by esophageal cancer Ec9706 cells was significantly higher than that of RGD / R8-LP, RGD-LP and LP, the difference was statistically significant (P <0.01). The cell viability of R8-LP-PTX, RGD-LP-PTX and LP- / R8-LP-PTX group 1.6 times, 1.7 times and 2.2 times, the difference was statistically significant (P <0.01). After 7 days of administration, the tumor spheres in the saline group continued to grow with a volume increase of 1.48 times. The volume of the tumor sphere in the LP-PTX group increased 1.12 times that of the original volume in the RGD / R8-LP-PTX group and the R8-LP-PTX group And tumor volume of RGD-LP-PTX group decreased to 36%, 59% and 64% of the original volume, the difference was statistically significant (P <0.01). CONCLUSION: Costimulatory RGD and cell-penetrating peptide R8 modified paclitaxel (PTX) liposomes can effectively penetrate the tumor cell membrane into the tumor cells, which is an effective drug delivery system for esophageal cancer chemotherapy.