Recombinant Newcastle disease virus rL-RVG induces ferroptosis in lung adenocarcinoma cells through the p53 YAP1-ACSL4 pathway
Objective The purpose of this study is to explore whether the recombinant Newcastle disease virus rL-RVG af-fects ferroptosis in lung adenocarcinoma cells through the p53 YAP1-ACSL4 pathway.Methods A549 and PC9 cell lines were cul-tured in vitro and cells in logarithmic proliferation were divided into control group(NC),Newcastle disease virus infection group(NDV),recombinant Newcastle disease virus infection group(rL-RVG),ferroptosis inducer group(Erastin)and ferroptosis inhibitor group(NAC).After viral infection and intervention with ferroptosis inducer and inhibitor,we determined the functional changes of cells,including cell vitality,migration ability,and invasion ability,using CCK-8,scratch experiments,and Transwell,and observed morphological changes in cells through optical microscopy.In addition,we used flow cytometry and fluorescence microscopy to deter-mine the content of ROS in cells,and used enzyme-linked immunosorbent assay to measure MDA content,and used Western blot and real-time PCR to detect the expression of ferroptosis related proteins p53,YAP1,and ACSL4.Results Compared to NC group,a significant decrease of cell growth,migration,and invasion of rL-RVG group was observed(P＜0.01).ROS and MDA levels were significantly augmented(P＜0.01).The levels of ROS and MDA in-creased significantly compared to the ferroptosis inducer group(P＜0.01),while the content of ferroptosis inhibitor decreased after inter-vention(P＜0.01).The expression of ferroptosis key proteins p53,YAP1,and ACSL4 were significantly augmented(P＜0.01).After knocking down YAP1 and ACSL4 by Si-RNA,the level of corresponding proteins were decreased(P＜0.01),and the content of ROS and MDA were also decreased(P＜0.01).Conclusion rL-RVG can effectively prevent the proliferation,migration,and diffusion of lung adenocarcinoma cells,and increase the content of lipid peroxides and cellular reactive oxygen species through the p53 YAP1-ACSL4 axis,ultimately promoting ferroptosis of tumor cells.
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