CPC/CMC-CD55sp纳米微球对宫颈癌Caski细胞的靶向杀伤作用  

Targeted antitumor effects of C-PC/CMC-CD55sp nanospheres on cervical carcinoma Caski cells

作　　者：刘慧慧 姬欢欢 朱峰 姜良乾 刘国祥 李冰 LIU Huihui;JI Huanhuan;ZHU Feng;JIANG Liangqian;LIU Guoxiang;LI Bing(Department of Biology, Faculty of Medicine, Qingdao University, Qingdao 266021, Shandong, China)

机构地区：[1]青岛大学医学部生物学教研室,山东青岛266021

出　　处：《中国肿瘤生物治疗杂志》2018年第12期1223-1229,共7页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81471546;81001346)~~

摘　　要：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.

关 键 词：藻蓝蛋白 羧甲基壳聚糖 CD55特异性配体肽 纳米微球 CASKI细胞 

分 类 号：R730.51[医药卫生—肿瘤] R737.33[医药卫生—临床医学]