Vafaei Sedigheh, Sadat Shandiz Seyed Ataollah, Piravar Zeinab
Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran.
Biol Trace Elem Res. 2020 Nov;198(1):109-117. doi: 10.1007/s12011-020-02054-6.
In the current study, zinc-phosphate nanoparticles (ZnPNPs) were investigated for the first time due to their anticancer activity against breast cancer Michigan Cancer Foundation-7 (MCF-7) cell line. The modification of such nanoparticles (NPs) was further examined for physicochemical characterization using various techniques such as powder X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential calculation, field emission scanning electron microscopy (FESEM), energy-dispersed spectroscopy (EDS), and Fourier transform infrared (FTIR) spectroscopy. Then, the newly fabricated ZnPNPs were tested for their in vitro cell cytotoxicity against breast cancer MCF-7 cells and noncancerous human embryonic kidney HEK293 cells, using MTT assay as a colorimetric one to assess cell metabolic activity for 24 h. The apoptotic efficacy of the NPs was subsequently confirmed through data obtained from Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining kit and cell cycle analysis. Determination of reactive oxygen species (ROS) generation was further performed via flow cytometry. Additionally, the expression of tumor suppressor genes p53 was analyzed using real-time polymerase chain reaction (PCR). Also, the prepared NPs showed a mean particle size of 38 nm. The measurements correspondingly showed that the cytotoxicity of MCF-7 cells depends on the concentration of NPs (IC = 80.112 μg/mL). MCF-7 cells were associated with initiation of apoptotic pathway in cells. Additionally, flow cytometry revealed cell cycle arrest in sub-G1 phase. ROS production was also obtained after treatment with IC concentration. According to annexin V-FITC/PI staining kit data, the percentage of early and late apoptotic cells was 78.2% in those treated with ZnPNPs. Moreover, the real-time PCR results demonstrated the ability of NPs in upregulating p53 gene expression. In summary, the data demonstrated that fabricated ZnPNPs had prominence to act as antitumor agents in breast cancer therapy.
在当前研究中，首次对磷酸锌纳米颗粒（ZnPNPs）进行了研究，因其对密歇根癌症基金会-7（MCF-7）乳腺癌细胞系具有抗癌活性。使用粉末X射线衍射（XRD）、动态光散射（DLS）、zeta电位计算、场发射扫描电子显微镜（FESEM）、能量色散光谱（EDS）和傅里叶变换红外（FTIR）光谱等各种技术，进一步研究了此类纳米颗粒（NPs）的改性，以进行物理化学表征。然后，使用MTT法作为比色法，测试新制备的ZnPNPs对乳腺癌MCF-7细胞和非癌性人胚肾HEK293细胞的体外细胞毒性，以评估细胞代谢活性24小时。随后，通过膜联蛋白V-异硫氰酸荧光素（FITC）/碘化丙啶（PI）染色试剂盒和细胞周期分析获得的数据，证实了NPs的凋亡效力。通过流式细胞术进一步测定活性氧（ROS）的产生。此外，使用实时聚合酶链反应（PCR）分析肿瘤抑制基因p53的表达。此外，制备的NPs的平均粒径为38nm。测量结果相应表明，MCF-7细胞的细胞毒性取决于NPs的浓度（IC = 80.112μg/mL）。MCF-7细胞与细胞凋亡途径的启动有关。此外，流式细胞术显示细胞周期停滞在亚G1期。用IC浓度处理后也获得了ROS产生。根据膜联蛋白V-FITC/PI染色试剂盒数据，用ZnPNPs处理的细胞中早期和晚期凋亡细胞的百分比为78.2%。此外，实时PCR结果表明NPs具有上调p53基因表达的能力。总之，数据表明制备的ZnPNPs在乳腺癌治疗中具有作为抗肿瘤剂的显著作用。
