Effect Of ShRNA Knockdown ECEL1 On Gene Expression Profile Of Liver Cancer Cells

Objective To investigate the effects of ECEL1 knockdown on gene expression profile of liver cancer cells.Methods1.Human liver cancer cells(BEL-7404)are divided into 2 groups,the experimental group(KD group): human liver cancer cells(BEL-7404)+ ECEL1 gene sh RNA lentivirus infection;control group(NC group): human liver cancer cells(BEL-7404)+ negative control virus infection.2.Extracted the total RNA in the Trizol method,purified the m RNA,and reversely transcribed the two groups of m RNA to synthesize fluorescently labeled c DNA mixture probes.3.Fluorescence-labeled c DNA mixture probe,hybridized with Gene Chip3'IVT Express Kit(Affymetrix),after strict washing,the scanner scans the chip fluorescence signal image,computer software data processing,and compared the obtained data to obtain the information about differences in gene expression between individuals.4.The screening criteria for differential genes were: | Fold Change |> 1.5 and FDR <0.05,P <0.05,further screening criteria for significant difference genes were: | Fold Change | ?2 and FDR <0.05,P <0.05,based on IPA,analysed classical signaling pathways,upstream regulation analysis,disease and function analysis,and regulation effect analysis were performed to clarify the changes of the interaction network.Results1.First,when the screening criteria were | Fold Change |> 1.5 and FDR <0.05,P <0.05,the gene chip technology was used to detect 748 differential genes expressed in the experimental group(KD group),371 genes were up-regulated and377 genes were down-regulated in the experimental group,and when the screening criteria were | Fold Change | ?2 and FDR <0.05,P <0.05,80 significant differential genes were further screened,of which the significance was significant,there were 34 up-regulated genes and 46 significantly down-regulated genes.2.Based on IPA analysis,compared with the control group(NC group),the experimental group(KD group)had 19 signal pathways,such as April Mediated Signaling and HGF Signaling,which were obviously activated,and the p38 MAPK Signaling signal pathway was significantly inhibited.16 upstream regulatory factors such as WNT3 A and TGFB1 were significantly up-regulated,and 30 upstream regulatory factors such as actinomycin D and STAT3 were significantly down-regulated,and a complex interaction network was formed.Conclusion1.Interfere with the expression of ECEL1 gene in human hepatoma cells(BEL-7404),and used gene chip technology to screen the effect of the knockdown of ECEL1 gene on the differentially expressed genes in human hepatoma cells(BEL-7404).2.Based on IPA,bioinformatics analysis was performed on the selected differential genes,and it was found that the knockdown of the ECEL1 gene had an effect on the classical signaling pathway,upstream regulatory factors,regulatory effects,diseases and functions of human hepatoma cells(BEL-7404),and formed a complex interaction network.3.The research provided an experimental basis for further elucidating the occurrence,development mechanism and therapeutic targets of primary liver cancer,and provided an experimental basis for further research.