Gene Screening And Expression Analysis Of Ameloblastoma Invasion Related Gene And Long Non-coding RNA

OBJACTIVE:Ameloblastoma is a kind of benign odontogenic tumor which is commonly found in oral and maxillofacial regions.Unlike general benign tumors,ameloblastoma has invasion biological features,which leads to a higher recurrence rate after surgery.lncRNA(long non-coding RNA)refers to non-coding RNA with a length of more than 200 bps,and is widely involved in the regulation of tumorigenesis,proliferation,invasion and metastasis.At present,there is no research on lncRNA in the process of invasion of ameloblastoma.The second generation of high-throughput total RNA sequencing is a technical means for detecting both mRNA and non-coding RNA.Screening and prediction of lncRNA and its target genes related to tumor regulation by high-throughput sequencing is a common research method in oncology research.METHODS:Collected 14 pairs samples of fresh ameloblastoma and its paraneoplastic tissues.Eight pairs of samples were sequenced using total RNA sequencing.Screening and prediction of lncRNA and genes related to ameloblastoma invasion by bioinformatics analysis.Real-time quantitative PCR(Real-time PCR)detection of the selected lncRNA and its target gene.Verify the analysis of changes in its expression.RESULTS:Through RNA sequencing,we screened a total of 110 pairs of lncRNA-target gene regulatoion combinations.Furthermore,through gene function analysis,we screened four combinations : LINC01007-COL26A1,LOC284080-ITGA3,LOC105378231-SPARC,and RPL21P101-ADAMTS20 which probably have lncRNA-target gene regulation functiong associated with ameloblastoma invasion.The relative expression of LINC01007 in ameloblastoma was 3.89 times higher than that in paraneoplastic tissues(P=0.024,t-test),and the relative expression of the corresponding target gene COL26A1 was 4.59 times(P>.=0.0024,t test);the relative expression of LOC284080 in ameloblastoma was 3.71 times that in paraneoplastic tissues(P=0.0018,ttest),and the relative expression level of target gene ITGA3 was 8.71 times(P= 0.0022,t test).There was no significant difference in the expression of LOC105378231 and PRL21P101 between ameloblastoma and paraneoplastic tissues(P>0.05.t test).At the same time,the expression of these two lncRNA target genes has obvious changes.The expression difference of SPARC was 1.98 times(P=0.0137,t-test),and the differential fold of expression of ADAMTS20 was 4.73-fold(P=0.00012,t-test).‘'CONCLUSION:In this study,total RNA sequencing,bioinformaticsl analysis and Real-time PCR were used to screen out lncRNAs that may be associated with ameloblastoma invasion: LINC01007,LOC284080,and genes: COL26A1,ITGA3,SPARC,ADAMTS20.It also suggests the possible regulatory mechanisms between LINC01007-COL26A1,LOC284080-ITGA3.We firsty report he function of these lncRNAs and target genes associated to invasion regulation in ameloblastoma,which can be the basement for the study about key regulation target of the invasion mechanism of ameloblastoma and can be the selection of targets in targeted therapeutic.