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Archive > Volume 28 Issue 8 > 2021,28(8):783-789. DOI:10.3872/j.issn.1007-385X.2021.08.003 Prev Next
Basic Research
Ultrasmall iron oxide nanoparticles inhibit the migration and invasion of human hepatocellular carcinoma HepG2 cells by enhancing autophagy
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Abstract:
Objective: To explore the effects of ultrasmall iron oxide nanoparticles (USIONPs) on the migration and invasion of human hepatocellular carcinoma HepG2 cells and its possible mechanism. Methods: The hydrate particle size and core particle size of USIONPs were analyzed by particle analysis device and transmission electron microscope, respectively. The dispersity and stability of USIONPs were characterized by Zeta potential and colloid stability analysis, respectively, to identify the successful prepaation of USIONPs. Different concentrations of USIONPs (0, 50, 100, 200 μg/ml) or 200 μg/ml USIONPs+5 mmol/L 3-MA (autophagy inhibitor) were used to treat human liver cancer HepG2 cells; then, CCK-8 assay was used to detect the proliferation viability of HepG2 cells; Transwell method was used to detect the migration and invasion ability of HepG2 cell; WB experiment was applied to detect the expression of autophagy markers Beclin1, LC3 and p62; 2',7'-dichlorodihydrofluorescein diacetic acid (DCFH-DA) method was used to determine the intracellular reactive oxygen species (ROS) level, and the iron ion colorimetric method was used to detect the iron ion level in the cells. Results: The average hydrate particle size of USIONPs was (37.86±12.90) nm, the core particle size was about 10 nm, and the Zeta potential was –23.8 mV, and confirmed the successful preparation of USIONP. The USIONPs had good water solubility and dispersibility. As the mass concentration of USIONPs increased and the incubation time prolonged, the proliferation viability of HepG2 cells showed a decreasing trend. Compared with the control group, after incubating HepG2 cells with 200 μg/ml USIONPs for 24 h, the numbers of migrated and invaded cells were significantly reduced (all P<0.05), and the addition of 3-MA could partially offset the above effects (P<0.05). Compared with the control group, the relative protein expression levels of beclin1 and LC3Ⅱin HepG2 cells in the 100 and 200 μg/ml USIONPs treatment groups increased significantly (all P<0.05), while the p62 protein expression decreased significantly (all P<0.05); 200 μg/ml USIONPs could significantly increase the level of intracellular ROS and iron ions, while the autophagy inhibitor 3-MA could block its effect (all P<0.05). Conclusion: USIONPs may promote autophagy of HepG2 cells, which consequently degrades USIONPs to release iron ions, leading to increased intracellular iron levels and ROS levels, thereby inhibiting the migration and invasion of HepG2 cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 32060228), the Natural Science Foundation of Guangxi Autonomous Region (No. 2017GXNSFAA198112, No. 2019GXNSFAA245077), the Postgraduate Education Innovation Project of Guangxi Autonomous Region (No. YCSW2019214, No. YCSW2020225), and the Scientific Research and Technology Development Project from Guilin City (No.20190219-2)
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