Pal Manish Kumar, Jaiswar Shyam Pyari, Dwivedi Ashish, Goyal Shruti, Dwivedi Vinay Nand, Pathak Anumesh Kumar, Kumar Vinod, Sankhwar Pushp Lata, Ray Ratan Singh
Department of Obstetrics and Gynecology, KGMU, Lucknow, 226003. India.
Department of Zoology, BHU, Varanasi. India.
Anticancer Agents Med Chem. 2017;17(12):1721-1732. doi: 10.2174/1871520617666170425094549.
Ovarian cancer is most lethal among all gynecologic malignancies. Paclitaxel (PTX) is well used chemotherapeutic regimen for cancer control; however its undesired toxicity has been a matter of concern for clinicians. Here, we used the graphene oxide coated nanotised apigenin (GO-NA) to enhance the efficacy of paclitaxel.
The combined use of paclitaxel (PTX) and nanotised apigenin (NA) may reduce the PTX dose and increase the efficacy.
GO and GO-Apigenin was prepared by modified Hummers method and the nanoparticles were characterized by dynamic light scattering and transmission electron microscopy. SKOV-3 cells were treated by DMSO, Group I (Control)-McCoy's 5A Medium, Group II-Paclitaxel (5nM), Group III- Nanotised Apigenin (GO-NA-10µM), Group IV- Paclitaxel (5nM) + GO-NA (10µM). Cell viability and IC-50 value were determined by MTT assay, synergism by Compusyn software, ROS by DCFH-DA assay, SOD activity by kit and MMP were examined by JC-1 and mitotracker/DAPI staining, cell cycle by flow cytometry, mRNA and protein level by Real Time-PCR and Western blot respectively Results: Results showed that GO-NA-PTX enhanced the anti-proliferative effect in synergistic manner as compare to GO-NA and PTX alone. GO-NA-PTX significantly suppressed the SOD activity, promotes the ROS accumulation, mitochondrial depolarization, DNA integrity and cell cycle arrest collectively accord the apoptosis. Results of immunocytochemistry, RT-PCR and western blot showed up-regulation of caspase-3, Bax, and down-regulation of Bcl-2.
The combination of PTX with GO-NA produces synergistic effects in SKOV-3 cells via the modulation of pro and anti-apoptotic gene and may reduce side effects of PTX.
卵巢癌是所有妇科恶性肿瘤中致死率最高的。紫杉醇（PTX）是用于癌症控制的常用化疗方案；然而，其不良毒性一直是临床医生关注的问题。在此，我们使用氧化石墨烯包覆的纳米化芹菜素（GO-NA）来提高紫杉醇的疗效。
联合使用紫杉醇（PTX）和纳米化芹菜素（NA）可能会降低PTX剂量并提高疗效。
采用改良的Hummers法制备氧化石墨烯和氧化石墨烯-芹菜素，通过动态光散射和透射电子显微镜对纳米颗粒进行表征。SKOV-3细胞分别用二甲基亚砜、I组（对照）- McCoy's 5A培养基、II组-紫杉醇（5nM）、III组-纳米化芹菜素（GO-NA-10µM）、IV组-紫杉醇（5nM）+ GO-NA（10µM）处理。通过MTT法测定细胞活力和IC-50值，使用Compusyn软件分析协同作用，通过DCFH-DA法检测活性氧，使用试剂盒检测超氧化物歧化酶活性，通过JC-1以及线粒体追踪染料/4',6-二脒基-2-苯基吲哚染色检测线粒体膜电位，通过流式细胞术检测细胞周期，分别通过实时定量PCR和蛋白质印迹法检测mRNA和蛋白质水平。结果：结果表明，与单独的GO-NA和PTX相比，GO-NA-PTX以协同方式增强了抗增殖作用。GO-NA-PTX显著抑制超氧化物歧化酶活性，促进活性氧积累、线粒体去极化、DNA完整性破坏和细胞周期停滞，共同导致细胞凋亡。免疫细胞化学、RT-PCR和蛋白质印迹结果显示，半胱天冬酶-3、Bax上调，Bcl-2下调。
PTX与GO-NA联合使用可通过调节促凋亡和抗凋亡基因在SKOV-3细胞中产生协同作用，并可能降低PTX的副作用。
