MiRNA And MRNA Expression Profiles And Docking Studies Of Oral Squamous Cell Carcinoma And Adjacent Tissue

[Background] oral squamous cell carcinoma is the highest incidence ofmalignant tumors in the head and neck cancer-prone cell invasion and metastasisresulting in poor prognosis. Oral squamous cell carcinoma in all aspects of clinicaldiagnosis, clinical treatment progress, but the survival rate of patients after fiveyears is still less than50%of this figure in the last30years has never been aneffective change. Studies have shown that miRNAs are key regulator of tumor celldevelopment, by matching the target gene protein-coding genes, inducing mRNAdegradation or translation inhibition, and to participate in the process of regulationof different tumor cell. The current study found that miRNAs play a key role in theoccurrence and development of oral squamous cell carcinoma, but their specificroles and mechanisms is not yet clear.The application of the subject HiSeq deep sequencing (high-throughput deepsequencing) method for detection of oral squamous cell carcinoma and pairedadjacent noncancerous tissue samples, with reference to the miRBase and Genebankdatabase, screening of differentially expressed miRNA and mRNA in oral squamouscell carcinoma (diff The expression of miRNA is defined as: Fold-change>1orFold-change <-1, and p-value <0.01; differentially expressed mRNA is defined as:the FDR≤0.001multiples of differences in2times more genes), build oralsquamous cell carcinoma difference between miRNA and mRNA expression profiling, to investigate the occurrence and development of expression variation inoral squamous cell carcinoma; from the expression level differences in miRNA andmRNA in oral squamous cell carcinoma; applications Real-time PCR method todetect mir-20a-5p, ODZ3and TCHH in oral squamous cell carcinoma and adjacenttissue of, validate, verify the reliability and accuracy of the sequencing data.[Objective] We apply HiSeq deep sequencing (high-throughput deep sequencing)method to construct miRNA and mRNA expression profiles, screening travelheterosexual expression of miRNA and mRNA; preliminary analysis of miRNA andmRNA differential expression of meaning in oral squamous cell carcinoma; anddifferentially expressed in oral squamous cell carcinoma of miRNA/mRNAdocking analysis. Theoretical and experimental basis for further study of the role ofmiRNA in oral squamous cell carcinoma and mechanisms.[Methods]10cases of surgical removal of fresh frozen next to the cancer of oralsquamous cell carcinoma and its paired normal tissues, Trizol one extraction of totalRNA spectrophotometer to measure the RNA concentration; application ofhigh-throughput sequencing technology to build oral squamous cell carcinomacancer and matched adjacent normal tissue of miRNA/mRNA expression profilesand analysis; by bioinformatics techniques to predict miRNA target genes; cancer ofthe oral squamous cell carcinoma and paired adjacent normal tissues significantdifferences of miRNA/mRNA expression profile docking analysis; applicationReal-Time PCR validation of differentially expressed miRNA and mRNAexpression in oral squamous cell carcinoma and adjacent tissues.[Results](1) Application HiSeq deep sequencing (high-throughput deep sequencing) methodand analyzed10oral squamous cell carcinoma and paired adjacent noncancerous tissues of miRNA expression, combined with data analysis, these differences ofmiRNA in oral squamous cell carcinoma Expression variation were analyzed,results showed that:77miRNAs were significantly differentially expressed,24miRNA downregulated,53miRNAs were up-regulated, suggesting that thesemiRNA and is closely related to the occurrence of oral squamous cell carcinoma.Test:19and14new miRNA in oral squamous cell carcinoma and adjacent tissuesare not registered in the miRBase database.(2) application HiSeq deep sequencing (high-throughput deep sequencing) methodto analyze the mRNA expression in oral squamous cell carcinoma and pairedadjacent noncancerous10pairs, combined with the data analysis of thesedifferences in mRNA in oral squamous cell carcinoma expression variation: pairingin oral squamous cell carcinoma and adjacent normal tissue were found14629mRNA in these,1120mRNA was significantly upregulated and178mRNAexpression down, suggesting that these mRNA was significantly differentiallyexpressed in oral squamous cell carcinoma is closely related.(3) by oral squamous cell carcinoma and paired adjacent tissues, significantdifferences of miRNA/mRNA expression profile docking analysis, we found that:different miRNA, the number of its docking target genes are not the same, there maybe zero, the number of possible thousands, for example: let-7c target genes5301;mir-17-5p target genes of6767;7614mir-19a-3p target genes; mir-20a-5p targetgenes of5865, the opposite is the mir-184, the target genes of mir-183-3p,mir-196a-5p and so indeed zero; differentially expressed genes in the dockingmiRNA large number of. By two or more miRNA regulation, such as: MAGEA11by let-7a-3p, let-7b-3p, let-7d-3p, let-7f-1-3p, let-7f-2-3p, mir-15a-5p, mir-16a-5p12miRNA regulation; COL1OA1mir-24-1-5p, mir-26a-5p, let-7a-3p, let-7b-3p,let-7d-3p11miRNA regulation; EMR1by the mir-17-3p, mir-19-3p, mir-19a-3p, mir-20a-3p, mir-25-3p, mir-26b-3p6miRNA regulation; this a similar mRNA alsoODZ3of, BAALC, LRRC17. The abnormal expression of these genes may berelated to the close regulation of the miRNA. Development of oral squamous cellcarcinoma, the regulation of differential gene expression of miRNA is not the same,their expression mechanism is not nearly the same, may be a miRNA-based,supplemented by to participate in a variety of miRNA.(4) Real-Time PCR experiments show that mir-20a-5p, ODZ3, TCHH expressionconsistent with the sequencing results, sequencing data is credible.[Conclusion] by the same batch of oral squamous cell carcinoma and pairedadjacent noncancerous tissue samples, significant differences in miRNA/mRNA indynamic expression profile studies and docking analysis initially revealed miRNAexpression variation of disorders in oral squamous cell carcinoma. further elucidatethe molecular mechanisms of oral squamous cell carcinoma of the development ofadequate theoretical basis for looking for an effective early diagnosis of oralsquamous cell carcinoma and malignant progression of biological molecularmarkers.