Research AURKA And SMAD3 Of 3'UTR By Alternative Cleavage And Polyadenylation In Liver Cancer

Background Alternative polyadenylation(APA)is mRNA select a particular kind of cleavage sites in the transcripts of maturation process,and then alternative polyadenylation.It is a widespread phenomenon and an important of control gene expression,in most eukaryotic gene containing multiple cleavage and polyadenylation sites,alternative cleavage and polyadenylation sites(pAs)can make the trancript develop an different mature mRNA.When pAs in 3'untranslated region(UTR),can lead to choose different pAs formed several mRNA isform.The mRNA isform is difference of the lost part from downstream 3'UTR after pAs.When pAs are alllocated in the 3'UTR,resulting in transcriptswith 3'UTRs of different length but encoding the same protein.A increasing number of examples illustrates that this phenomenon can significantly influence the expression of gene,and plays an important role in mRNA metabolism,APA also plays a important role in the pathogenesis of cancer.Objective APA is thought to be associated with the occurrence and development of the many human diseases.In the 3'UTR of AURKA and SMAD3 may be have alternative cleavage and polyadenylation sites,this pAs can make AURKA and SMAD3 produce different length of mRNA isform.In recent years,many studies have shown that APA pattern shortening of 3'UTR by alternative cleavage and polydenylation in cancer,and related with the occurrence and development of the tumor,This study is to investigate AURKA and SMAD3 gene produce different abundance mRNA isform for APA,the relationship between primary hepatocellular carcinoma and APA,also hope APA patterns help to us looking for novel biomarkers provide valuable clues.Method Retrievethe National Center for Biotechnology Information(NCBI)before the experiment started,download HCC sample datasets GSE12941?GSE19665 in Gene Expression Omnibus(GEO)publication dates.We have the using high-throughput technology for detection of gene expression profiles in 38 pairs of liver tissue and adjacent of cancer,then get exon array data sets.Using bioinformatics method(probelevelalternative transcript analysis,PLATA)to GSE12941?GSE19665 and exon array data sets carry out alternative splicing identification,then select candidate genes AURKA and SMAD3 of success.Based on AURKA and SMAD3 mRNA isform have different lengths sequence features,two pairs of primers were designed.Normal liver cell line and HEPG2 cell line for RNA extraction,by RT-PCR(reverse transcription PCR)technology for reverse transcription of RNA,then the PCR product for sanger sequencing.With semi-quantitative reverse transcription and polymerase Chain reaction(SqRT-PCR)technique to examination AURKA and SAMD3 expression level in 38 pairs of liver tissue and adjacent of cancer.Results AURKA and SMAD3 are all have pAs located in the 3'UTR.AURKA and SMAD3 in L02 cell line.HEPG 2 cell line?liver tissue and adjacent of cancer have confirmed the existence of two different mature mRNA isform.AURKA with shorting 3'UTR expression level was not statistical difference in liver tissue and adjacent of cancer(P>0.05).But SMAD3 with shorting 3'UTR expression level was statistical difference in liver tissue and adjacent of cancer(P<0.05)Conclusion AUEKA and SMAD3 exists pAs can make gene happened alternative cleavage and polyadenylation,AURKA with shorting 3'UTR was not statistical difference in liver tissue,SMAD3 with shorting 3'UTR was statistical difference in liver tissue.For the occurrence and development of HCC provides a new mechanism.