Roy Kislay, Patel Yogesh S, Kanwar Rupinder K, Rajkhowa Rangam, Wang Xungai, Kanwar Jagat R
Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), Centre for Molecular and Medical Research (C-MMR), School of Medicine (SoM), Faculty of Health, Deakin University, Waurn Ponds, VIC, Australia.
Institute for Frontier Materials (IFM), Deakin University, Waurn Ponds, VIC, Australia.
Int J Nanomedicine. 2015 Dec 21;11:25-44. doi: 10.2147/IJN.S91810. eCollection 2016.
This study used the Eri silk nanoparticles (NPs) for delivering apo-bovine lactoferrin (Apo-bLf) (~2% iron saturated) and Fe-bLf (100% iron saturated) in MDA-MB-231 and MCF-7 breast cancer cell lines. Apo-bLf and Fe-bLf-loaded Eri silk NPs with sizes between 200 and 300 nm (±10 nm) showed a significant internalization within 4 hours in MDA-MB-231 cells when compared to MCF-7 cells. The ex vivo loop assay with chitosan-coated Fe-bLf-loaded silk NPs was able to substantiate its future use in oral administration and showed the maximum absorption within 24 hours by ileum. Both Apo-bLf and Fe-bLf induced increase in expression of low-density lipoprotein receptor-related protein 1 and lactoferrin receptor in epidermal growth factor (EGFR)-positive MDA-MB-231 cells, while transferrin receptor (TfR) and TfR2 in MCF-7 cells facilitated the receptor-mediated endocytosis of NPs. Controlled and sustained release of both bLf from silk NPs was shown to induce more cancer-specific cytotoxicity in MDA-MB-231 and MCF-7 cells compared to normal MCF-10A cells. Due to higher degree of internalization, the extent of cytotoxicity and apoptosis was significantly higher in MDA-MB-231 (EGFR+) cells when compared to MCF-7 (EGFR-) cells. The expression of a prominent anticancer target, survivin, was found to be downregulated at both gene and protein levels. Taken together, all the observations suggest the potential use of Eri silk NPs as a delivery vehicle for an anti-cancer milk protein, and indicate bLf for the treatment of breast cancer.
本研究使用 Eri 蚕丝纳米颗粒(NPs)在 MDA-MB-231 和 MCF-7 乳腺癌细胞系中递送载脂蛋白牛乳铁蛋白(Apo-bLf)(约 2%铁饱和)和铁饱和牛乳铁蛋白(Fe-bLf)(100%铁饱和)。与 MCF-7 细胞相比,尺寸在 200 至 300 nm(±10 nm)之间的载有 Apo-bLf 和 Fe-bLf 的 Eri 蚕丝 NPs 在 MDA-MB-231 细胞中 4 小时内显示出显著的内化。用壳聚糖包被的载有 Fe-bLf 的蚕丝 NPs 进行的体外肠袢试验能够证实其在口服给药方面的未来用途,并显示在 24 小时内回肠的吸收量最大。Apo-bLf 和 Fe-bLf 均诱导表皮生长因子(EGFR)阳性的 MDA-MB-231 细胞中低密度脂蛋白受体相关蛋白 1 和乳铁蛋白受体的表达增加,而 MCF-7 细胞中的转铁蛋白受体(TfR)和 TfR2 促进了 NPs 的受体介导的内吞作用。与正常 MCF-10A 细胞相比,从蚕丝 NPs 中可控且持续释放的两种 bLf 在 MDA-MB-231 和 MCF-7 细胞中均显示出更强的癌症特异性细胞毒性。由于内化程度较高,与 MCF-7(EGFR-)细胞相比,MDA-MB-231(EGFR+)细胞中的细胞毒性和凋亡程度明显更高。发现一种突出的抗癌靶点 survivin 的表达在基因和蛋白质水平均下调。综上所述,所有观察结果表明 Eri 蚕丝 NPs 作为抗癌乳蛋白递送载体的潜在用途,并表明 bLf 可用于治疗乳腺癌。