Wnt3、Wnt3a Influence On Apoptosis Of Hepatic Progenitor Cell

ObjectiveThe use of adenovirus Wnt3, Wnt3a transfected liver precursor cells,flow cytometry method marked by Annexin-PE/7-ADD and Hoechst33342staining detected apoptosis status, Real-time PCR and Western blot analysisexpression of apoptotic and anti-apoptotic factors. Learn overexpression ofWnt3, Wnt3a impact on apoptosis of hepatic progenitor cells.Methodsliver precursor cells Transfecte Wnt3, Wnt3a gene, the detection ofWnt3, Wnt3a gene：①Real-time Quantitative PCR was used to detectmRNA expression of Wnt3, Wnt3a;②Western Blotting assay Wnt3, Wnt3aprotein expression levels.Wnt3, Wnt3a in liver progenitor cells apoptosis:①Real-time PCRand immunobiotting detect the expression levels of Bcl-2, Bax apoptoticgenes such as nucleic acids and proteins;②Flow Cytometry assayapoptosis rate;③Hoechst33342staining cell assay apoptosis.ResultWnt3, Wnt3a gene was upregulated in the liver precursorcells,Adenovirus containing Wnt3, Wnt3a gene successfully transfected hepatic progenitor cells. After transfection of liver cell precursors,Hoechst33342staining showed pyknotic nuclei of the characteristicchanges，it reduce the number of apoptotic; Flow Cytometry showed that theapoptosis rate decreased; Real-time PCR and immunobiotting test resultsthat anti-apoptotic Bcl-2and other gene was highly expressed in mRNA andprotein, the expression of pro-apoptotic factor Bax reduced.ConclusionThis study showed that Wnt3、Wnt3a in overexpression of liverprecursor cells inhibits apoptosis, the mechanism may be work throughBcl-2family.