目的 制备肝癌细胞膜嵌合的TPP修饰HCPT脂质体（HCPT@T/HM-L），将HCPT高效递送到肝癌细胞线粒体，充分发挥药物在靶细胞器的药效作用。方法 采用薄膜水化法制备HCPT@T/HM-L，以挤压法将肝癌细胞膜嵌合在脂质体膜上，测定粒径、ζ电位、电镜进行表征；然后检测体外释放、溶血；以香豆素6为荧光探针，探索其胞内转运、线粒体靶向效果；最后从肿瘤细胞和荷瘤小鼠模型上，系统性评价肝癌细胞膜嵌合的HCPT@T/HM-L抗肿瘤效果。结果 HCPT@T/ HM-L在电镜下呈现为规则圆球、具有“核-壳”状结构，粒径大约为（162.7±5.8）nm，ζ电位为（12.4±1.8）mV；HCPT@T/HM-L能促进载药系统的细胞摄取和靶向进入线粒体；细胞药效结果显示，HCPT@T/HM-L能很大程度上抑制肝癌细胞的生长，降低线粒体膜电位、升高细胞内ROS和Caspase-3水平，降低抗凋亡蛋白Bcl-2，提高促凋亡蛋白Bax表达，从而增强药物促进肝癌细胞的大量凋亡；荷瘤小鼠结果显示，HCPT@T/HM-L能明显抑制皮下瘤的生长，促使肿瘤细胞线粒体出现严重的功能障碍，从而促进肿瘤细胞的大量凋亡；细胞和动物试验结果均显示，HCPT@T/HM-L具有很好的抗肿瘤效果，明显优于其他组，这可能与肝癌细胞膜的融合以及TPP阳离子的线粒体靶向作用有关。结论 HCPT@T/HM-L可以高效递药到肝癌细胞线粒体，促进肿瘤细胞的大量凋亡，在精确治疗肿瘤方面具有较好的临床转化潜能。

Objective To prepare TPP-modified 10-hydroxycamptothecin liposomes (HCPT@T/HM-L) embedded in liver cancer cell membranes for the efficient delivery of hydroxycamptothecin to mitochondria of hepatocellular carcinoma cells to give full play to the efficacy of the drug in the target organella. Methods TPP-modified 10-hydroxycamptothecin liposomes were prepared by thin film hydration method, and liver cancer cell membranes were embedded on liposome membranes using the extrusion method, and characterized by particle size, ζ potential and electron microscopy; Then in vitro release and hemolysis were detected; Coumarin 6 was used as a fluorescent probe to explore its intracellular transport and mitochondrial targeting effects; Finally, the anti-tumor effects of chimeric TPP-modified 10-hydroxycamptothecin liposomes were systematically evaluated in tumor cells and tumor-bearing mice. Results HCPT@T/HM-L exhibited regular spheres with a “core-shell” structure under electron microscopy. The particle size was approximately (162.7 ± 5.8) nm, and the ζ potential was (12.4 ± 1.8) mV. The efficacy results showed that HCPT@T/HM-L largely inhibited the growth of hepatocellular carcinoma cells, decreased mitochondrial membrane potential, increased intracellular ROS and Caspase-3 levels, decreased anti-apoptotic protein Bcl-2, and increased the expression of pro-apoptotic protein Bax, thus enhancing the promotion of massive apoptosis of hepatocellular carcinoma cells by the drug; the results in ruminant mice showed that HCPT@T/HM-L significantly inhibited the growth of subcutaneous tumors and induced severe mitochondrial dysfunction in tumor cells, thus promoting massive apoptosis of tumor cells; Both cellular and animal test results showed that HCPT@T/HM-L had a great anti-tumor effect, which was significantly better than other groups, which might be related to the fusion of liver cancer cell membranes and the mitochondrial targeting effect of TPP cations. Conclusion HCPT@T/HM-L could efficiently deliver drugs to the mitochondria of hepatocellular carcinoma cells and promote massive apoptosis of tumor cells, which has good clinical translational potential in the precise treatment of tumors.

R283.6

2020—2021年度河南省中医药科学研究专项课题（20-21ZY1019）