MIR-494 Inhibits Tumor Progression By Targeting LETMD1 In Cervical Cancer

Objective:Cervical cancer is one of the most common female reproductive system malignant tumors.Due to changes in people's lifestyles and other factors,the morbidity of cervical cancer has been increasing in recent years.LETMD1 is also known as the human cervical cancer gene?HCCR?.A large number of studies have shown that LETMD1 is related to the development of tumors and it plays an important role in tumor markers.As an oncogene,LETMD1has been proved overexpressed in both cervical cancer and cervical cancer cells,but the specific mechanism remains unclear.MicroRNA?miRNA?are small non-coding RNA with a length of about 22 nucleotides.They are widely found in eukaryotes and play a significant role in a variety of physiological and pathological processes.MiR-494 was first found in human retinoblastoma.At present,more and more studies have proved that miR-494is widely involved in the occurrence and development of malignant tumors.The potential mechanism of miR–494 needs to be further explored although few studies have reported the effect of miR–494 on the occurrence and development in cervical cancer.In this study,bioinformatics technology,RT-qPCR,western blot,dual-luciferase reporter assay and cell function assay were used to explore the role of miR-494 in the pathogenesis of cervical cancer through targeted LETMD1.Methods:1.MiR-494 and LETMD1 protein expression were determined by RT–qPCR and western blot in 30 clinical cervical cancer tissues and 30normal cervical tissues respectively to analyze their relationship and their relationship with the clinicopathological features of patients.2.miR-494 mimic,miR-494 inhibitor and their negative controls were transfected into HeLa cells using Lipofectamine²⁰⁰⁰ to explore the expressions of miR-494 and LETMD1 in the transfected cells.3.Bioinformatics technology was used to predict the targeting relationship between miR-494 and LETMD1,and dual-luciferase reporter assay experiment was used to confirm the targeted regulation between miR-494 and LETMD1.4.Furthermore,the effects of overexpression and low expression of miR-494 on the proliferation of HeLa cells were detected by clonal formation assay.Transwell assay was further used to detect the effects of overexpression and low expression of miR-494 on invasion and migration of HeLa cells.All data were analyzed by one-way analysis of variance?ANOVA?using SPSS 22.0 software.P<0.05 was considered to be statistically significant.Results:1.Compared with normal cervical tissues,miR-494 expression was significantly decreased in cervical cancer tissues,and the expression of LETMD1 was significantly increased in cervical cancer tissues.Interestingly,LETMD1 and miR-494 levels exhibit negative correlation?P<0.05?,and miR-494 expression level and the FIGO stage also showed negative correlation?P<0.05?.2.Compared with the mimic NC,LETMD1 mRNA and protein levels were significantly decreased when transfected with miR-494 mimic?P<0.05?.while compared with the inhibitor NC,LETMD1 mRNA and protein levels were significantly increased when transfected with miR-494 inhibitor?P<0.05?.3.Bioinformatics predicted that LETMD1 was the target gene of miR-494,and dual-luciferase reporter assay further confirmed that miR-494directly acted on the 3'UTR of LETMD1 mRNA.4.The results of clonal formation assay showed that overexpression of miR-494 could inhibit the proliferation of HeLa cells?P<0.05?,while Transwell assay confirmed that overexpression of miR-494 could significantly inhibit the migration and invasion of HeLa cells?P<0.05?.Conclusion:The expression of miR-494 was significantly correlated with the FIGO stage of patients,and miR-494 could inhibited the proliferation,migration and invasion of cervical cancer cells by targeting LETMD1.