The Effect of Over-Expressing FGD6 on the Differentiation in Hepatic Stem Cells
YU Shishi1, SHA Ou1, LI Wanling1, HU Qian1, HE Tongchuan2, ZHANG Bingqiang1*
The aim of this study was to investigate the effect of over-expressing FGD6 (faciogenital dysplasia 6) on the differentiation of hepatic stem cells and its possible mechanism. The FGD6 gene was inserted into an adenovirus vector to over-express and packaged into an adenovirus (Ad-FGD6) to infect hepatic stem cells HP14.5, where the empty vector adenovirus group (Ad-null group) and the uninfected adenovirus group (Blank control group) were used as comparisons. The mRNA and protein expression levels of FGD6, hepatic stem cell marker (AFP), hepatocyte markers (ALB), bile duct epithelial cell markers (CK19 and SOX9) and the canonical Wnt pathway markers (β-catenin and wnt3a) were detected by semi-quantitative PCR and Western blot respectively. The change in intracellular glycogen synthesis was detected by glycogen staining (PAS staining), and the cell proliferation was detected by CCK8. The results showed that the mRNA and protein levels of FGD6 when HP14.5 cells were infected by the Ad-FGD6 adenovirus were increased compared with the Ad-null group and the Blank control group (P<0.05). The mRNA and protein levels of hepatic stem cell marker (AFP) and bile duct epithelial markers (CK19, SOX9) were decreased (P<0.05). But the mRNA and protein levels of hepatocyte marker (ALB) and the canonical Wnt pathway markers were elevated (P<0.05). There were the purplish-red granules in the Ad-FGD6 group that represented the positive reaction, which indicated that the HP14.5 had the function of glycogen synthesis. CCK8 detection found that Ad-FGD6 group can promote hepatic stem cell proliferation (P<0.05). Therefore, the up-regulation of FGD6 gene expression can facilitate HP14.5 cells to differentiate into hepatocytes and promote cell proliferation, which may be achieved through the canonical Wnt pathway.
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