Expression And Function Of MiR-1293-KCNAB2 Axis In Lung Adenocarcinoma

BACKGROUND: Lung cancer is the number one cancer with the highest death rate not only in China but also worldwide.Although some risk factors(e.g.,smoking,air pollution,etc.)have been shown to be associated with the development of lung cancer,scientists have not been able to fully explain how lung cancer is a major contributor to human health.Therefore,identification of molecular markers associated with lung cancer progression is essential for both early diagnosis and the development of effective treatment plans.miRNAs are a hot area of research recently,and they are present in most tumorigenesis and metastasis,and there are also many studies proving their involvement in the biological activities of lung adenocarcinoma development.The miR-1293 studied in this project is currently a relatively novel miRNA and has not yet been studied.This trial aims to investigate the effect of miR-1293 on cell phenotype in lung adenocarcinoma and further explore its target KCNAB2,which opens up new ideas for gene targeting therapy for lung cancer.AIM: To explore the biological function and target of miR-1293 in the occurrence and development of lung adenocarcinoma.METHODS: 1.The original miRNAs date of lung adenocarcinoma were downloaded from The Cancer Genome Atlas(TCGA)database,and the micro RNAs with distinct differential expression were screened,and the heat map and volcano maps were made.The expression of miR-1293 in lung adenocarcinoma was confirmed by UALCAN website,and the relationship between miR-1293 and prognosis of lung adenocarcinoma was explored.mir-1293’s potential target genes were predicted by the bioinformatics method,and then the target genes were enriched for functional analysis on David website.Secondly,according to the combined score ranking on Targetscan,the first 9target genes were selected from these target genes,and finally the target gene KCNAB2 was selected.To explore the relationship between the expression of KCNAB2 and the prognosis of patients with lung adenocarcinoma using GEPIA database,three pairs of specimens of lung adenocarcinoma and adjacent tissues were used for immunohistochemistry to verify the expression of KCNAB2.2.The mimics and inhibitors of miRNA-1293 were transiently transfected into A549 cell line and the efficiency of transfection mimics was verified by real-time quantitative PCR.Ed U assay explored the effect of miR-1293 on proliferation ability of A549 cells,Transwell migration assay explored the effect of miR-1293 on cell migration ability.Phagokinetic motility assay explored the effect of miR-1293 on cell motility,Transwell invasion assay explored the effect of miR-1293 on cell invasion ability.western blot method examined the effect of miR-1293 on the change of KCNAB2 protein expression.Dual-luciferase assay was performed to verify that KCNAB2 is a target gene of miR-1293.RESULTS:1.miR-1293 was highly expressed in lung adenocarcinoma tissues(TCGA,UALCAN),and its expression was negatively correlated with its prognosis.2.KCNAB2 was lowly expressed in lung adenocarcinoma tissues(GEPIA),its expression was positively correlated with prognosis,and it was verified by immunohistochemistry that KCNAB2 expression was reduced in lung adenocarcinoma.3.miR-1293 mimic promoted the proliferation,migration and invasion of A549 cells.The opposite result could be verified when the expression of miR-1293 was inhibited.4.Biological information methods indicated that KCNAB2 may be the direct target of miR-1293.The binding site was verified by dual-luciferase experiment,and the negative correlation between miR-1293 and KCNAB2 protein expression was verified by western blotting.CONCLUSION:1.The high expression of miR-1293 in lung adenocarcinoma promotes the proliferation,migration and invasion of lung adenocarcinoma A549.2.KCNAB2 is low expressed in lung adenocarcinoma and is negatively regulated by miR-1293.