Vatani Mahsa, Hosseinzadeh Simzar, Sari Amirhossein, Ghomi Marzdashti Hamidreza, Rahimpour Azam, Fattahi Roya
Plasma Physics Research Center, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Pharm Res. 2024 Nov 12;23(1):e150385. doi: 10.5812/ijpr-150385. eCollection 2024 Jan-Dec.
Invasive cervical cancer is recognized as the second most common malignancy in women after breast cancer.
This study investigates, for the first time, the effect of gold nanoparticle-doped graphene oxide (GO) nanosheets on the human epithelial carcinoma (HeLa) cell line in the presence of heliox cold plasma.
Graphene oxide nanosheets were synthesized using the Hummer method and then doped with gold nanoparticles. The nanoparticles were characterized by transmission electron microscopy (TEM), and the diffraction peaks of GO and gold nanoparticles were confirmed through X-ray diffraction (XRD) analysis. Additionally, the optical absorbance of the nanoparticles was measured in the range of 200 - 900 nm using UV-Visible spectroscopy. A plasma generator was fabricated to produce cold plasma using helium (He) and oxygen (O₂) gases at a 99:1 ratio. The radicals generated by the cold plasma were analyzed via optical emission spectroscopy (OES). Cell treatment was conducted by applying various concentrations of GO and GO/Au nanoparticles. Cellular phenotype was monitored through optical microscopy, and biocompatible concentrations of both nanoparticles were determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Subsequently, cold plasma at varying distances and durations was applied to the nanoparticle-treated cells. The generated radicals and the expression of apoptotic genes in treated cells were assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and real-time PCR, respectively.
The width of the bacillus-like gold nanoparticles was 15.13 ± 0.96 nm. The cold plasma generated radicals such as N2I2⁺, N2II1⁻, He•, and O⁻•. XRD analysis confirmed the successful coupling of gold onto the GO nanosheets. The biocompatible concentrations of GO and GO/Au nanoparticles were found to be 30 µg/100 µL and 20 µg/100 µL, respectively, as determined by the MTT assay. Radical formation increased as incubation time was extended from 30 to 60 seconds. Furthermore, real-time PCR analysis demonstrated the highest levels of p53, Bax, and caspase 3/8 expression at a plasma exposure time of 60 seconds in the composite-treated group, while Bcl2 expression was significantly reduced.
The findings suggest that the parameters of heliox cold plasma and the concentrations of GO/Au nanoparticles must be optimized to effectively induce apoptosis in cervical cancer cells.
浸润性宫颈癌被认为是女性仅次于乳腺癌的第二大常见恶性肿瘤。
本研究首次探究了在氦氧冷等离子体存在的情况下，金纳米颗粒掺杂的氧化石墨烯（GO）纳米片对人上皮癌细胞系（HeLa）的影响。
采用Hummer法合成氧化石墨烯纳米片，然后用金纳米颗粒进行掺杂。通过透射电子显微镜（TEM）对纳米颗粒进行表征，并通过X射线衍射（XRD）分析确认GO和金纳米颗粒的衍射峰。此外，使用紫外可见光谱在200 - 900 nm范围内测量纳米颗粒的吸光度。制造了一台等离子体发生器，以99:1的比例使用氦气（He）和氧气（O₂）产生冷等离子体。通过光发射光谱（OES）分析冷等离子体产生的自由基。通过施加不同浓度的GO和GO/Au纳米颗粒进行细胞处理。通过光学显微镜监测细胞表型，并使用3 - [4,5 - 二甲基噻唑 - 2 - 基] - 2,5 - 二苯基四氮唑溴盐（MTT）法测定两种纳米颗粒的生物相容性浓度。随后，对经纳米颗粒处理的细胞施加不同距离和持续时间的冷等离子体。分别使用2,2 - 二苯基 - 1 - 苦基肼（DPPH）和实时PCR评估处理后细胞中产生的自由基和凋亡基因的表达。
杆菌状金纳米颗粒的宽度为15.13±0.96 nm。冷等离子体产生了诸如N2I2⁺、N2II1⁻、He•和O⁻•等自由基。XRD分析证实金成功地耦合到了GO纳米片上。通过MTT法测定，GO和GO/Au纳米颗粒的生物相容性浓度分别为30 µg/100 µL和20 µg/100 µL。随着孵育时间从30秒延长到60秒，自由基形成增加。此外，实时PCR分析表明，在复合处理组中，血浆暴露时间为60秒时，p53、Bax和caspase 3/8的表达水平最高，而Bcl2表达显著降低。
研究结果表明，必须优化氦氧冷等离子体参数和GO/Au纳米颗粒浓度，以有效诱导宫颈癌细胞凋亡。
