Ultrasonic Assisted Cerium Oxide/Graphene Oxide Hybrid: Preparation, Anti-proliferative, Apoptotic Induction and G2/M Cell Cycle Arrest in HeLa Cell Lines

In this work, we developed a morphology involved cerium oxide/graphene oxide hybrid (CeO2/GO hybrid) nanocomposite based system using the ultrasonic method. X-Ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM) have been performed to analyse the developed nanocomposite. The crystallite size of the hybrid was found to be 3.86 nm comparably lower than that of pure CeO2 (7.4 nm) which was confirmed by X-Ray diffraction analysis. The presence of functional groups was affirmed using FTIR analysis. The morphological features of pure CeO2 and hybrid were verified using FESEM. FESEM images reveals that, pure CeO2 nanoparticles are highly agglomerated and CeO2/GO hybrid possess nano-rectangular morphology. In addition, the developed hybrid system has been analysed to evaluate its scavenging anti-cancer potential against HeLa cell lines at various concentrations and incubation intervals using antiproliferative assay test. The test results reveals that, as the concentration of hybrid nanocomposite increases the cell death also increases. Here, “IC50” refers to lowest concentration of sample (CeO2/GO hybrid) at which nearing 50% of HeLa cells remains alive and rest of the cells remains dead. In our study, 31.2 µg/ml is considered to be IC50 value for which nearing 50% of cells remains dead and it has been attained at an incubation period of 72 h. An dual acridine orange/ethidium bromide (AO/EB) fluorescent staining, was performed at two specific concentrations (i.e.) 125 µg/ml and 1000 µg/ml of developed hybrid nanocomposite to identify apoptosis-associated changes of cell membranes during the process of apoptosis. Change in color for both live cells (green) and dead cells (red) were viewed using fluorescence microscopy. The quantitative analysis was done using flow cytometry (FACS study) to investigate the cell cycle at which maximum number of HeLa cells has been killed due to interaction with developed CeO2/GO hybrid. The FACS study test results reveals that, maximum cancer cells were arrested at R3 (G2/M) phase. Hence, the developed CeO2/GO hybrid has shown improved anticancer efficacy against HeLa cell line and thus it acts as a better therapeutic agent for cervical cancer diagnosis.