The Mechanism Of MiR-34c-3p Regulating Ferroptosis In Oral Squamous Cell Carcinoma

Objective: Oral squamous cell carcinoma(OSCC)is one of the most common malignant tumors of the head and neck,but the five-year survival rate is low.MiRNA is a small molecule RNA that plays an important role in a variety of physiological and pathological processes.,and miR-34c-3p has been demonstrated to be closely related to the occurrence of tumors.Ferroptosis is a new type of cell death characterized by lipid peroxidation of cell membrane and organelle membrane.However,it is still unclear how miR-34c-3p influences the development of oral squamous cell carcinoma(OSCC)by regulating ferroptosis.Therefore,the main objective of this study was to explore the role and mechanism of miR-34c-3p in OSCC,to provide a new target for clinical diagnosis and treatment.Methods: 1.The expression of miR-34c-3p in OSCC tissues was lower than that in normal tissues,and was significantly correlated with tumor size,lymph node metastasis,long-term prognosis and other clinical characteristics.It was also confirmed that the expression of miR-34c-3p was significantly higher in OSCC cells than in normal cells.2.Cell models with knockdown and overexpression of miR-34c-3p were constructed by transfection of miR-34c-3p inhibitor and miR-34c-3p mimics,and the effects of miR-34c-3p on OSCC cell proliferation were detected by CCK-8,clonogenesis assay,live/dead cell staining,and flow cytometry.3.The possible downstream target genes of miR-34c-3p were analyzed by bioinformatics,and the correlation was verified by double luciferase assay,immunohistochemical analysis,q RT-PCR and western blot.4.Whether the target gene is involved in Mir-34C-3p regulation of OSCC cell proliferation was verified by cloning formation experiment,CCK-8 experiment,live/dead cell staining experiment,flow cytometry and other functional recovery experiments.5.Reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)analysis,Western Blot detection of glutathione peroxidase 4(GPX4)and CCK-8 detection of cell viability after adding apoptosis inhibitors and iron death inhibitors.To verify whether miR-34c-3p regulates iron death in OSCC cells through target genes.Results: 1.miR-34c-3p was lower expressed in OSCC tissues than in paracancer tissues,and was significantly correlated with tumor size,lymph node metastasis,long-term prognosis and other clinical features.The results were verified by comparing tumor cells with normal cells.2.miR-34c-3p inhibitor and mimics can effectively knock down or overexpress miR-34c-3p.Compared with the negative control group,overexpression of miR-34c-3p can significantly inhibit the proliferation of SCC-25 cells and promote cell death.3.The target gene of miR-34c-3p was determined by Target Scan database analysis,and SLC7A11 was confirmed as the direct target gene of miR-34c-3p by double luciferase assay,q RT-PCR,Western blot and other further experiments.4.Overexpression of miR-34c-3p in SCC-25 cells can significantly inhibit cell proliferation,and overexpression of SLC7A11 plasmid SLC7A11 can reverse the inhibition of miR-34c-3p on SCC-25 cells.5.Overexpression of miR-34c-3p can promote cell iron death,and co-transfection of overexpressed SLC7A11 plasmid can remove the promoting effect of miR-34c-3p on cell iron death.Conclusion: Our findings revealed a novel strategy to upregulate erastin-induced ferroptosis in OSCC through the miR-34c-3p/SLC7A11 axis,suggesting new insights into OSCC and a potentially useful therapeutic strategy for OSCC.Conclusions: Our results indicate that miR-34c-3p is expressed lowly in tumors,revealing a novel strategy for inhibiting OSCC proliferation by up-regulating erastin-induced iron death by miR-34c-3p /SLC7A11 axis,and providing a potential therapeutic strategy for the diagnosis and treatment of OSCC.