The Effect And Mechanism Of Ferroptosis Induced By FCRA NPs In Oral Squamous Cell Carcinoma Cells

Introduction:Background:Oral squamous cell carcinoma(OSCC)is the most common malignant tumor of Oral epithelial origin in the maxillofacial region,accounting for 2%to 4%of all tumors and more than 80%of head and neck tumors.The prognosis is poor,with a 5-year survival rate of less than 50%.Chemotherapy is one of the main methods for the treatment of oral cancer,but there are many limitations in the current chemotherapy drugs,such as poor selectivity,lack of precision targeting,large adverse reactions,easy to produce tumor resistance,etc.Therefore,it is of great significance to study the precise targeted treatment of tumor.This experiment application of Ferrocene retinoic acid nanoparticles(FCRA NPs),by ferrocene modified retinoic acid molecules self-assembly,and mainly consists of retinoic acid and Ferrocene two functional groups.Previous experiments have shown that retinoic acid functional groups of FCRA NPs are of a similar structure and the activity of retinoic acid can induce tumor stem cells,ferrocene functional groups of main control FCRA Nps aggregation and dissociation,and it is sensitive to oxidative reducing,the tumor cells in high content of reduced Glutathione(GSH)sensitive,after the FCRA Nps encounter with GSH,immediate disintegrate,thus play a role of effective targeting.The content of GSH in normal cells is low,and the nanoparticle will not or rarely dissociate,thus reducing the toxic and side effects on normal tissues and cells.The ferrocene part of FCRA NPS depletes GSH in tumor cells and increases intracellular Reactive oxygen species(ROS),which are highly similar to the biochemical characteristics of iron death.Iron death is oxidative cell death induced by small molecules,which is an iron-dependent regulatory cell death triggered by accumulation of lipid peroxides.When tumor cells develop drug resistance,different small molecule substances can directly or indirectly act on GPX4 to induce iron death of cells,which can play a strong inhibitory effect on tumor growth,that is,improve the sensitivity of tumor cells to chemotherapy drugs.Therefore,inducing iron death of cells plays an important role in reversing drug resistance of tumors and improving therapeutic effect.Therefore,this study will use FCRA NPS to act on oral squamous cell carcinoma cells,and study the role of FCRA NPS in inducing iron death of oral squamous cell carcinoma cells,regulatory pathways and key targets,so as to provide a theoretical basis for the development of drugs or nanomaterials targeting iron death and its key regulatory targets,and further explore more optimal treatment of oral squamous cell carcinoma.Purpose:To investigate the role,regulatory pathways and key targets of iron death induced by FCRA NPS in oral squamous cell carcinoma cells,and to provide a theoretical basis for improving the therapeutic effect of oral cancer.MethodsFirstly,human oral squamous cell carcinoma cells were cultured to study the inhibitory effect of FCRA NPS on the proliferation,migration and invasion of oral squamous cell carcinoma cells,so as to clarify the inhibitory effect of FCRA NPS on oral squamous cell carcinoma cells.Further with FCRA Nps oral squamous cancer cells after processing,through a cellular level(GSH,ROS)testing,subcellular level(mitochondrial morphology),morphology(transmission electron microscope cell morphology)and molecular level(iron death related protein GPX4,SLC7A11 transcription and expression)and so on several aspects of iron death characteristics,ultimately determine the FCRA Nps iron death induced by oral squamous cancer cells and its mechanism.ResμLts:(1)FCRA NPs can inhibit the growth,migration and invasion of oral squamous cell carcinoma cells.(2)FCRA NPs acted on squamous cell carcinoma cells,intracellular GSH decreased,ROS aggregation,It was preliminarily confirmed that FCRA NPS induced ferroptosis of oral squamous cell carcinoma cells.(3)Mitochondria fluorescence staining showed that the mitochondria were smaller and the nuclear size did not change significantly.(4)The morphology of the cells under transmission electron microscope showed that the mitochondria were smaller,the membrane density was increased,the mitochondrial ridges were reduced or disappeared,the outer membrane of the mitochondria was broken,the nucleus size was normal,but the chromatin aggregation was lacking.(5)The changes of P53,Nrf2,Slc7a11 and Gpx4 were detected by Q-PCR,and the mechanism of Ferroptosis induced by FCRA NPs in oral squamous cell carcinoma cells was preliminarily determined.Conclusion:FCRA NPs can induce iron death of oral squamous cell carcinoma cells.FCRA NPs inhibits the expression of SLC7A11 by acting on key targets such as P53 and Nrf2,resulting in the failure of cystine uptake by System XC~-on the cell membrane to synthesize GSH,which affects the activity of GPX4,leads to accumulation of lipid peroxides and promotes iron death of cells.