陆军军医大学学报 (Jul 2024)
Recombinant Newcastle disease virus rL-RVG induces ferroptosis of gastric cancer cells through inhibiting Nrf2-GCLC-GPX4 pathway
Abstract
Objective To investigate whether recombinant Newcastle disease virus (rL-RVG) induces iron death in gastric cancer cells through Nrf2-GCLC-GPX4 pathway. Methods After human gastric cancer HGC-27 cells were treated with rL-RVG, Newcastle disease virus (NDV) and PBS solution (control group), respectively, cell proliferation, invasion and migration were detected by CCK-8 assay and Transwell invasion assay and cell scratch test. Ferroptosis accelerator (erastin), and nuclear factor E2 related factor 2 (Nrf2) accelerator (TBHQ) and inhibitor (ML385) were added respectively as controls. The content of malondialdehyde (MDA) in each treatment group was detected by lipid oxidation kit. The content of reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe and flow cytometry. Western blotting and immunofluorescence assay were employed to measure the expression levels of Nrf2-GCLC-GPX4 pathway related proteins. Results Compared with the control group, the survival rate of HGC-27 cells were significantly decreased after rL-RVG and NDV treatment in a dose- and time-dependent manner, and the effect was more significant in the rL-RVG treatment group (P < 0.05). The migration and invasion abilities of HGC-27 cells were obviously inhibited in the NDV and rL-RVG treatment groups, and the latter had more notable inhibition than the former. The protein levels of Nrf2, GCLC, SLC7A11 and GPX4 were statistically decreased (P < 0.05), and the contents of MDA and ROS were increased (P < 0.05) in the virus treatment groups than the control group, with the increasing or decreasing trend more significant in the rL-RVG group than the NDV group. What's more, the protein levels of SLC7A11 and GPX4 were decreased in the erastin group (P < 0.05). Compared with the control group, those of Nrf2, GCLC, SLC7A11 and GPX4 were increased in the TBHQ group and decreased in the ML385 group (P < 0.05), while the contents of MDA and ROS were decreased and increased respectively in the above 2 groups (P < 0.05). Compared with the rL-RVG group, the rL-RVG+TBHQ group and rL-RVG+ML385 group had enhanced and reduced protein expressio of Nrf2, GCLC, SLC7A11 and GPX4, respectively (P < 0.05), while the contents of MDA and ROS were in opposite trends (P < 0.05). Conclusion rL-RVG can induce ferroptosis of gastric cancer cells through Nrf2-GCLC-GPX4 pathway, and then inhibit the growth of tumor cells.
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