Liu Guoxiang, Xu Xiaohui, Jiang Liangqian, Ji Huanhuan, Zhu Feng, Jin Bingnan, Han Jingjing, Dong Xiaolei, Yang Fanghao, Li Bing
Department of Genetics and Cell Biology, Basic Medical College, Qingdao University, Qingdao, China.
Department of Medical Genetics, Linyi People's Hospital, Linyi, China.
Front Pharmacol. 2020 Jun 18;11:906. doi: 10.3389/fphar.2020.00906. eCollection 2020.
studies had shown that C-Phycocyanin (C-PC) inhibited cervical cancer HeLa cells growth. We constructed C-PC/CMC-CD55sp nanospheres using C-PC, Carboxymethyl Chitosan (CMC), and CD55 ligand peptide (CD55sp) to allow for targeted antitumor effects against HeLa cells and . The characteristics of the nanospheres were determined using FTIR, electron microscopy, and laser particle size analysis. Flow cytometry, laser confocal microscopy and small animal imaging system showed the targeting of C-PC/CMC-CD55sp nanospheres on HeLa cells. Subsequently, the proliferation and apoptosis were analyzed by Cell Counting Kit-8 (CCK-8), flow cytometry, TUNEL assay and electron microscopy. The expression of the apoptosis-related protein was determined using western blot. The stainings of Hematoxylin and Eosin (HE) were employed to evaluate the cell condition of tumor tissue sections. The cytokines in the blood in tumor-bearing nude mice was determined using ELISA. These results showed that C-PC/CMC-CD55sp nanospheres were successfully constructed and targeted HeLa cells. The constructed nanospheres were more effective than C-PC alone in inhibiting the proliferation and inducing apoptosis in HeLa cells. We also found that C-PC/CMC-CD55sp nanospheres had a significant inhibitory effect on the expression of antiapoptotic protein Bcl-2 and a promotion on the transformation of caspase 3 to cleaved caspase 3. C-PC/CMC-CD55sp nanospheres played an important role in tumor suppression, reduced the expression TGF-β, and increased IL-6 and TNF-α. This study demonstrates that the constructed new C-PC/CMC-CD55sp nanospheres exerted targeted antitumor effects and which provided a novel idea for application of C-PC, and provided experimental basis for comprehensive targeted treatment of tumors.
研究表明，C-藻蓝蛋白（C-PC）可抑制宫颈癌HeLa细胞的生长。我们使用C-PC、羧甲基壳聚糖（CMC）和CD55配体肽（CD55sp）构建了C-PC/CMC-CD55sp纳米球，以实现对HeLa细胞的靶向抗肿瘤作用。通过傅里叶变换红外光谱（FTIR）、电子显微镜和激光粒度分析来确定纳米球的特性。流式细胞术、激光共聚焦显微镜和小动物成像系统显示了C-PC/CMC-CD55sp纳米球对HeLa细胞的靶向作用。随后，通过细胞计数试剂盒-8（CCK-8）、流式细胞术、TUNEL检测和电子显微镜分析细胞增殖和凋亡情况。使用蛋白质免疫印迹法测定凋亡相关蛋白的表达。采用苏木精和伊红（HE）染色来评估肿瘤组织切片的细胞状况。使用酶联免疫吸附测定（ELISA）法测定荷瘤裸鼠血液中的细胞因子。这些结果表明，C-PC/CMC-CD55sp纳米球成功构建并靶向HeLa细胞。所构建的纳米球在抑制HeLa细胞增殖和诱导凋亡方面比单独的C-PC更有效。我们还发现，C-PC/CMC-CD55sp纳米球对抗凋亡蛋白Bcl-2的表达有显著抑制作用，并促进半胱天冬酶3向裂解的半胱天冬酶3转化。C-PC/CMC-CD55sp纳米球在肿瘤抑制中发挥重要作用，降低转化生长因子-β（TGF-β）的表达，并增加白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）。本研究表明，所构建的新型C-PC/CMC-CD55sp纳米球具有靶向抗肿瘤作用，为C-PC的应用提供了新思路，并为肿瘤的综合靶向治疗提供了实验依据。
