目的:制备一种新型C-藻蓝蛋白（C-phycocyanin,CPC）/羧甲基壳聚糖-CD55 配体肽（carboxymethyl chitosan-CD55-specific ligand peptide, CMC-CD55sp）(CPC/CMC-CD55sp)纳米微球，并探究其对宫颈癌Caski 细胞的靶向治疗作用。方法：采用离子交联法制备新型CPC/CMC-CD55sp 纳米微球，通过透射电镜（DLS）和红外光谱仪（FTIR）观察纳米微球的表征，Western blotting和流式细胞术检测CD55 在Caski 细胞和成纤维（L-929）细胞表面的表达情况，CCK-8 法检测纳米微球对Caski 细胞增殖的影响，流式细胞术和荧光显微镜检测纳米微球被细胞摄取情况,Western blotting 和流式细胞术检测纳米微球对Caski 细胞凋亡相关信号蛋白和凋亡率的影响,溶血试验测定药物的生物安全性。结果：成功制备新型CPC/CMC-CD55sp 纳米微球，其形态良好、直径分布均匀，CD55 在宫颈癌细胞Caski 表面高表达而在小鼠L-929 细胞表面低表达（P<0.01）。纳米微球能靶向、高效地到达Caski细胞并被摄入细胞；其在人外周血中溶血作用微弱，且在安全范围；其对Caski 细胞的增殖具有明显的抑制作用并且能够诱导Caski 细胞凋亡（P<0.05 或P<0.01）。结论：新型CPC/CMC-CD55sp 纳米微球能够靶向抑制宫颈癌Caski 细胞的增殖并诱导其凋亡，具有较好的安全性，为海洋抗肿瘤药物的研发提供了新思路。

Objective: To prepare a new type of phycocyanin / carboxymethyl chitosan-CD55 ligand peptide (CPC /CMC-CD55sp)nanospheres, and to study its targeted therapeutic effect on cervical cancer Caski cells. Methods: The novel CPC/CMC-CD55sp nanospheres (CPC/CMC-CD55sp) were synthesized by ionic cross-linking method, and the properties of nanospheres were observed by transmission electron microscopy (DLS) and fourier transform infrared spectroscopy (FTIR). The expression of CD55 on the surface of Caski and fibroblast (L-929) cells was detected by Western blotting and flow cytometry. The effect of nanospheres on the proliferation of Caski cells was detected by CCK-8. Flow cytometry and fluorescence microscopy were used to detect the uptake of microspheres by Caski cells; Western blotting and flow cytometry were used to detect the effect of CPC/CMC-CD55sp on expressions of apoptosis-related proteins and apoptosis rate in Caski cells; the hemolysis test was used to determine the biological safety of the drug. Results: CPC/CMC-CD55sp was successfully prepared with good morphology and uniform diameter; and CD55 was highly expressed on the surface of Caski cells but low expressed on the surface of L-929 cells (P<0.01). CPC/CMC-CD55sp could targeted and efficiently reach Caski cells and be ingested into the cells. It exhibited weak hemolysis effect on human peripheral blood, which was in the safe range. CPC/CMC-CD55sp displayed obvious inhibitory effect on Caski cell proliferation, and could induce cell apoptosis (P<0.05 or P<0.01). Conclusion:The new CPC/CMC-CD55sp can targeted inhibit the growth of cervical cancer Caski cells via inducing its apoptosis and has good bio-safety, which provides a new idea for the research and development of anti-tumor marine drugs.

国家自然科学基金资助项目（No.81471546, 81001346)