The Role Of METTL14-mediated M6a Methylation Of LncRNA NEAT1 In The Progression And Metastasis Of Renal Cancer

Background and objectiveLong Non-coding RNA(Lnc RNA)has become a new gene regulator and prognostic marker for many kinds of cancers.Lnc RNA nuclear rich transcriptional 1(NEAT1)is associated with tumorigenesis.It has been reported that the expression of NEAT1 is up-regulated in renal clear cell carcinoma(Clear cell renal cell carcinoma,cc RCC).In patients with renal cell carcinoma,the high expression of NEAT1 is related to tumor progression and poor survival.NEAT1 gene knockout inhibits the proliferation of renal cancer cells and inhibits the migration and invasion of renal cancer cells by reversing the phenotype of epithelial-to-mesenchymal transformation.At present,more than 100 kinds of RNA modifications have been found in organisms.Some studies have widely reported that some types of RNA modified nucleated m RNA,include N1-methyladenosine(m1A),N6-methyladenosine(m6A)and 5-methylcytosine(m5C),of which m6 A was first discovered in the 1970 s.At present,the transcriptional modification of m6 A epitopes has attracted people's attention.m6 ARNA modification is the most abundant internal modification of m RNA and non-coding RNA in most eukaryotes.The physiological importance of m6 ARNA modification has been confirmed by its key role in tissue development and differentiation.m6 A RNA modification regulates m RNA,such as structure,maturation,stability,splicing,output,translation and decay at different levels.In addition,m6 A RNA modification is also involved in cell fate determination,cell cycle regulation,cell differentiation and maintenance of circadian rhythm,and a large number of RNA binding protein(RBPs)is also affected by m6 A RNA modification.m6 A RNA modification is a dynamic and reversible posttranscriptional modification process maintained by a multicomponent methyltransferase �writer' complex(METTL3,METTL14 and their cofactors such as WTAP,etc)and removed by demethylase �eraser'(FTO and ALKBH5).The function of m 6 A in m RNA metabolism primarily depends on reader proteins,which include YT521-B homology(YTH)domain family(YTHDF1-3,YTHDC1-2).Although m6 A modification of RNA has been found to be associated with different types of cancer,little is known about the relationship between m6 A related genes and cc RCC.Therefore,from the relationship between RNAm6 A modification related genes and NEAT1 molecules,we explore the effect and mechanism of m6 A modification related genes on NEAT1,which may provide new clues for the treatment of renal cell carcinoma.Methods1.To detect the expression of METTL14 in renal cell carcinoma and the effect of METTL14 on the proliferation and migration of renal cell line,and verify that METTL14 can affect the effect of NEAT1 on the function of renal cell carcinoma.2.According to the NEAT1 sequence information,the existence of m6 A modification site in NEAT1 was inferred,and the interaction between METTL14 and NEAT1 in RNA m6 A writers was explored.3.Explore the combination of NEAT1 and m6 A reader YTHDF2,to verify the combination of NEAT1 and YTHDF2.4.To explore the interaction between NEAT1 expression and YTHDF2.5.To detect the expression characteristics and functional effects of YTHDF2 in renal cell carcinoma,and to verify the effect of YTHDF2 on the function of NEAT1 in renal cell carcinoma.6.To verify the relationship between NEAT1 and YTHDF2 and its effect on clinical prognosis by clinical samples.Results1.METTL14 is low expressed in renal cell carcinoma and inhibits the proliferation and migration of renal cell carcinoma.It can also affect the tumor-promoting effect of NEAT1 in renal cancer cells.2.Through the m6 A modification site prediction software SRAMP,it was predicted that there were m6 A modification sites on NEAT1,which were negatively regulated by METTL14 and had a binding relationship with METTL14.3.YTHDF2 can bind to NEAT1,and reduce the expression of NEAT1.4.The low expression of YTHDF2 in renal cell carcinoma inhibits the proliferation and migration of renal cancer cells,and affects the function of renal cancer cells by inhibiting the expression of NEAT1.5.The expression of NEAT1 was negatively correlated with the prognosis of the patients,and the expression of YTHDF2 was negatively correlated with the expression of NEAT1.ConclusionMETTL14 and YTHDF2 affect the expression and function of NEAT1 through m6 A.METTL14 and YTHDF2 play a role in inhibiting tumor proliferation and migration in renal cell carcinoma.m6 A modification of NEAT1 is expected to be a new target for gene targeting therapy of renal cell carcinoma.