Construction of stably transfected STATb-RNAi in HepG2 and identification of its function

Objective To explore the effect of RNAi lentivirus vector on stability HepG2 cell proliferation,migration. Methods The virus infected the HepG2 cells stably were divided into four groups: control group( untreatment),negative control group( infected with PLV-control vector),pLVTHmSTAT6-RNAi group,and pLVTHm-STAT6-ORF goup. Expressing STAT6-ORF and STAT6-RNAi cell lines stably were obtained,and the STAT6 expression were detected by PCR and Western blot. CCK-8 assay and Transwell assay were used to detect the stably transfected cell lines activity and cell migration ability changes. Results The STAT6-ORF and STAT6-RNAi HepG2 stable expression cell line were obtained successfully by TET screening.Compared with control cells,cell proliferation in STAT6-ORF overexpression group were fast( F = 4. 279,P = 0. 021),and had high mobility; and in STAT6-RNAi group,the cell proliferated slowly,with the culture time prolonged and passage to the cells of the third generation( HepG2 most cells have died),the cells gradually disintegrated,floating die,cell activity decreased significantly( F = 290. 114,P 0. 001),transfer capacity decreased significantly. Conclusion The STAT6-ORF and STAT6-RNAi HepG2 stable expression cell line were obtained successfully,is convenient to establish the animal model of late,tumor formation in nude mice,biological functions in vivo validation of STAT6,confirme the clinical significance of the gene.