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Optimized preparation of novel C-phycocyanin-carboxymethyl chitosan nanoparticles and its inhibitory effects on human cervical carcinoma HeLa cells proliferation
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Abstract:
Objective: To develop a methodology of preparing novel C-phycocyanin-carboxymethyl chitosan nanoparticles(C-PC/CMCNPs) and determine the effect of C-PC/CMCNPs on the growth of HeLa cells. Methods: An orthogonal experiment was designed with the particle diameter and entrapment efficiency as index and CMC: C-PC mass ratio, CMC concentration, and CaCl2 concentration as factors to determine the best preparing condition of C-PC/CMCNPs. The effects of the generated C-PC/CMCNPs on the growth and apoptosis of HeLa cells were assessed by CCK-8 assay and flow cytometry respectively. Caspase-3 protein expression in HeLa cells was quantified by Western blotting. Results: The optimal condition for C-PC/CMCNPs preparations were as follows: C-PC to CMC ratio of 1∶2, CMC concentration of 1 mg/ml and CaCl2 concentration of 1 mg/ml. The C-PC/CMCNPs prepared in these optimal conditions had a high entrapment efficiency with an average particle diameter of 118.4 nm. C-PC, CMC and C-PC/CMCNPs were all capable of inhibiting proliferation and inducing apoptosis in HeLa cells by up-regulating the expression of the caspase-3 protein, but the effect of C-PC/CMCNPs was significantly more pronounced (P<0.05). Conclusions: We have optimized the conditions of preparing C-PC/CMCNPs. The nanoparticles prepared under these conditions have an acceptable safety profile of sustained C-PC release and possess a caspase-3-dependent anti-tumor activity, suggesting potential clinical implications.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No. 81001346，No.81471546），the Medical Health Science and Technology Development Plan of Shandong Province (No. 2011HZ023)，and the Science and Technology Support Plan in Public Sphere from the Science and Technology Bureau of Qingdao City （No.2012-1-3-5-(4)-nsh）
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