Medical Biotechnology Department, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Technology Applications (SRTA-City), New Borg EL-Arab, 21934, Alexandria, Egypt.
Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Technology Applications (SRTA-City), New Borg EL-Arab, 21934, Alexandria, Egypt.
Sci Rep. 2017 Dec 1;7(1):16769. doi: 10.1038/s41598-017-16962-6.
Bovine lactoperoxidase (LPO) and lactoferrin (LF) are suitable proteins to be loaded or adsorbed to chitosan nanoparticles (NPs) for preparing stable nanoformulations with potent anticancer activity. In the present study, nanocombinations of LPO and LF revealed improvement in their stability and activity compared to single (free or nanoformulated) bovine proteins. The coating or loading of LPO-loaded NPs with LF resulted in the highest synergistic cytotoxicity effect against Caco-2, HepG-2, MCF-7 and PC-3 cells in comparison with other NPs and free proteins without causing toxicity toward normal cells. This synergistic improvement in the anticancer activity was apoptosis-dependent that was confirmed by severe alterations in cellular morphology, high percentage of annexin-stained cells and sub-G1 populations as well as nuclear staining with orange fluorescence of treated cancer cells. Additionally, significant alterations in the expression of well characterized cellular proliferation and apoptosis guards (NF-κB, Bcl-2 and p53) in these NPs-treated cancer cells compared to 5-fluorouracil (5-FU) treated cells. Our findings provide for the first time that these new synergistic nanoformulated forms of LPO and LF were superior in their selective apoptosis-mediating anticancer effect than free form of these proteins and 5-FU. LF coating or loading of LPO-loaded NPs present as promising therapy for cancer.
牛乳铁蛋白（LF）和乳过氧化物酶（LPO）是适合被负载或吸附到壳聚糖纳米粒子（NPs）上的蛋白质，可用于制备具有强大抗癌活性的稳定纳米制剂。在本研究中，与游离或纳米制剂的牛蛋白相比，LPO 和 LF 的纳米组合显示出稳定性和活性的提高。用 LF 对负载 LPO 的 NPs 进行涂层或负载，与其他 NPs 和游离蛋白相比，对 Caco-2、HepG-2、MCF-7 和 PC-3 细胞产生了最高的协同细胞毒性作用，而对正常细胞没有毒性。这种抗癌活性的协同增强是凋亡依赖性的，这通过细胞形态的严重改变、 Annexin 染色细胞和亚 G1 群体的高百分比以及用橙色荧光染色的处理癌细胞的核证实。此外，与 5-氟尿嘧啶（5-FU）处理的细胞相比，这些 NPs 处理的癌细胞中，特征明确的细胞增殖和凋亡保护因子（NF-κB、Bcl-2 和 p53）的表达发生了显著改变。我们的研究结果首次表明，与游离形式的这些蛋白质和 5-FU 相比，这些新的协同纳米制剂形式的 LPO 和 LF 在介导选择性细胞凋亡的抗癌作用方面更具优势。LF 对负载 LPO 的 NPs 的涂层或负载可能是癌症的一种有前途的治疗方法。
