The Effects And Mechanism Analysis Of Long Non-coding LncRNA SFTA1P In Huh7 Cells

Objective:1.Using bioinformatics technique to detected the differential expression of long non-coding RNA(lncRNAs)between hepatic carcinoma and para-cancerous tissues.2.To investigate the effects of lncRNA SFTA1 P on the biological function of human hepatoma cell line Huh7.3.Using Transcriptome sequencing(RNA-Seq)methods predicts the potential targets of SFTA1 P,and uses q RT-PCR technology to verify the targets.Method:1.Download the original file of human hepatic carcinoma data from the TCGA database,use the R language to analyze the data,and define the significantly up-regulated SFTA1 P as the research object of this experiment,and analysis the basic information of SFTA1 P through the bioinformatics website.2.Using Quantitative Real-time PCR(q RT-PCR)technology to verify the expression of SFTA1 P in hepatic carcinoma cells and human normal liver cells LO2.3.Using RNA interference technology and gene overexpression technology to down-regulate or up-regulate the expression level of SFTA1 P in Huh7 cells.4.The effects of SFTA1 P on proliferation of Huh7 cells were evaluated by CCK8 assay,The effects of SFTA1 P on migration of Huh7 cells were observed by scratch test,and the effects of SFTA1 P on apoptosis of Huh7 cells were detected by Flow cytometry assay.5.RNA-Seq technology analysis the differentially expressed mRNA in Huh7 cells after knocking down SFTA1 P,and predict the potential targets of SFTA1 P based on biological information.6.Using qRT-PCR technology to verify related differentially expressed genes.Result:1.Through bioinformatics analysis,it is found that SFTA1 P is up-regulated in hepatic carcinoma and is a pseudogene-derived lncRNA.2.The expression level of SFTA1 P in Huh7 cells was significantly up-regulated compared with LO2 cells.3.Compared with the control group,knocking down SFTA1 P can inhibit the proliferation and migration of Huh7 cells and promote cell apoptosis,while Up-regulate SFTA1 P can promote the proliferation and migration of Huh7 cells and inhibit cell apoptosis.4.RNA-Seq detected 393 differential mRNAs after knocking down SFTA1 P in Huh7 cells,of which 267 were up-regulated and 126 were down-regulated.5.Knockdown of SFTA1 P can up-regulate the transcription of DUSP1 and JUNB genes in Huh7 cells,while down-regulate the SFTA1 P can inhibit the transcription of DUSP1 and JUNB genes in Huh7 cells.Conclusion:lncRNA SFTA1 P may negatively regulate the expression of tumor suppressor factors DUSP1 and JUNB,thereby promoting the proliferation and migration of Huh7 cells and inhibiting cell apoptosis.