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目的制备整合素受体精氨酸—甘氨酸—天冬氨酸(Arg—Gly—Asp,RGD)和细胞穿膜肽(transcriptional actirator protein,TAT共修饰载紫杉醇(paclitaxel,PTX)脂质体(liposomes,LP)(RGD/TAT—LP—PTX),对其理化性质进行表征,并考察脂质体与肺癌A 549细胞的亲和力以及对A 549细胞增殖的抑制作用。方法采用薄膜分散法制备RGD/TAT—LP—PTX,考察脂质体的粒径,电位以及包封率,通过定量细胞摄取实验考察肺癌A 549细胞对RGDD/TAT—LP的摄取效率。构建A 549肿瘤球模型,定性共聚焦实验观察肿瘤细胞和肿瘤球对脂质体的摄取。噻唑蓝实验考察RSD/TAT-LP—PTX对肺癌A 549细胞的增殖抑制作用。结果 RGD/TAT—LP—PTX的粒径在(118.5±11.4)nm,电位为(21.58+2.42)mV,紫杉醇的包封率为86.5%。细胞摄取实验结果显示,PGD/TAT-LP在4 h摄取效率是2 h的1.88倍,差异具有统计学意义(P<0.05);肺癌A 549细胞在与脂质体共同孵育4 h后对RGD/TAT-LP的摄取效率分别是TAT—LP、RGD—LP和LP的2.1倍、2.8倍和5.1倍,差异具有统计学意义(P<0.01);MTT实验结果显示,在相同给药浓度下,不同脂质体组间细胞存活率有显著性差异(P<0.01)。结论整合素受体RGD和细胞穿膜肽TAT共修饰载紫杉醇(PTX)脂质体能够有效识别高表达整合索受体的肿瘤细胞,穿透肿瘤细胞膜进入肿瘤细胞,是一种潜在高效的肿瘤靶向给药系统。
OBJECTIVE: To study the effects of Arg-Gly-Asp (RGD) and the transcriptional actirator protein (TAT) on liposomes (PTX) , LP) (RGD / TAT-LP-PTX) were used to characterize their physico-chemical properties and to investigate the affinity of liposomes to A 549 cells and to inhibit the proliferation of A 549 cells.Methods The RGD / TAT-LP-PTX, the particle size, potential and entrapment efficiency of liposomes were investigated, and the uptake efficiency of RGDD / TAT-LP in lung cancer A 549 cells was investigated by quantitative cell uptake assay.A 549 tumor spheroid model, The uptake of liposomes by tumor cells and tumor spheres was observed experimentally.The inhibitory effect of RSD / TAT-LP-PTX on the proliferation of lung cancer A 549 cells was investigated by MTT assay.Results The particle size of RGD / TAT-LP-PTX was (118.5 ± 11.4) nm, the potential was (21.58 + 2.42) mV and the entrapment efficiency of paclitaxel was 86.5%. The results of cell uptake showed that the uptake efficiency of PGD / TAT-LP at 4 h was 1.88 times of that at 2 h, the difference was statistically significant (P <0.05). After lung cancer A 549 cells incubated with liposomes for 4 h, RGD / TAT-LP The uptake efficiency was 2.1 times, 2.8 times and 5.1 times higher than that of TAT-LP, RGD-LP and LP, respectively (P <0.01). The results of MTT assay showed that under the same drug concentration, (P <0.01) .Conclusion The coexpression of integrin receptor RGD and cell-penetrating peptide TAT modified paclitaxel (PTX) liposomes can effectively identify tumor cells with high expression of integrin receptor, Through the tumor cell membrane into the tumor cells, is a potentially efficient tumor targeting delivery system.