To Establish A Lentiviral Expression System Of CXCL-1and The Role Of CXCL-1in Hepatocyte

Objective The study found the chemokine CXCL-1and its receptor canpromote the proliferation of various cells. But the relationship between the trendof reported development of cytokines and their receptors and liver disease islittle, therefore, this study tries to construct CXCL-1lentiviral expressionsystem,to explore the correlation of chemokine CXCL-1and normal liver cells,and laythe foundation for further study of hepatocellular carcinoma cells.Methods Objective to construct the CXCL-1lentiviral expression system,screening the normal liver cell line of carrying GFP fluorescence labeling andover-expression of chemokine CXCL-1. Expanding culture, CXCL-1geneexpression in liver cells transfected identified by Western blot. Based on theestablished cell lines, through MTT assay and colony formation assay to studythe CXCL-1proliferation ability of normal liver cells in vitro and by usingTranswell experimental detection of target gene CXCL-1on normal liver cellmigration. The data from the experiment using SPSS16.0software package fordata analysis using repeated measures ANOVA comparing before and aftertransfection of normal liver cell lines (MTT experiments, colony formationassay,flow cytometry cell cycle experiments, Transwell experiments),whether a significant difference P<0.05was considered statistically significant.Results1)CXCL-1gene lentiviral expression system was constructedsuccessfully, virus titer was2.00E+8TU/ml;2)After transfection of48h by flowcytometry, three plasmid proportion is4:3:2transfection rate was the highest upto about95%;3) In the CXCL-1/7702group, clone formation and the number ofclone formation rate is higher than that of the control group (two thenonparametric Wilcoxon test shows that P<0.05);4)MTT assay results showedthat in the CXCL-1/7702group, the liver cell growth speed is faster than emptyvector group and blank control group;5)Transwell experiment showed: normalliver cell line7702has no migration,CXCL-1had no effect on7702normal livercell line transfer ability.Conclusion Chemokine CXCL-1has obvious cell growth promoting functionon normal liver cells, but no promote malignant transform function on normalliver cell.