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Chang C.-F.,Henan Normal University | Chang C.-F.,Henan Engineering Laboratory for Bioengineering and Drug Development | Zhao W.-M.,Henan Normal University | Zhao W.-M.,Henan Engineering Laboratory for Bioengineering and Drug Development | And 10 more authors.
Acta Anatomica Sinica | Year: 2016

Objective: To explore the effect of dexamethasone on cell proliferation and apoptosis in the rat liver cell line BRL-3A in vitro. Methods: BRL-3A cells were treated with different concentrations of dexamethasone, and MTT method was used to observe the effect of dexamethasone on cell activity at 12, 24, 48, 72 and 120 hours after treatment. Annexin V-FITC staining and propidium iodide (PI) staining were used to detect the effect of cell apoptosis and cell cycle. Real-time PCR was used to evaluate the changes in the expression of related genes. Results: The result of MTT assays revealed that dexamethasone inhibited the proliferation of BRL-3A cells in a dose-dependent manner. Annexin V-FITC staining showed that dexamethasone significantly induced the apoptosis of BRL-3A. PI staining indicated that the ability of proliferation decreased in the cells treated with dexamethasone. Real-time PCR analysis showed that pro-apoptosis genes Caspase-3, Caspase-8 and Caspase-9 were up-regiilated, while pro-proliferation genes Ccndl and Jun were down-regulated. Conclusion: Dexamethasone may inhibit the proliferation of BRL-3A cell line and induce its apoptosis by up-regulating Caspase-3, Caspase-8 and Caspase-9 and down-regulating the expression of Ccndl and Jun.

Ding Y.,Henan Normal University | Ding Y.,Henan Engineering Laboratory for Bioengineering and Drug Development | Chang C.,Henan Normal University | Chang C.,Henan Engineering Laboratory for Bioengineering and Drug Development | And 12 more authors.
Cytotechnology | Year: 2016

Hepatocytes differentiated from induced pluripotent stem cells and adult stem cells could be utilized as a tool for the study of liver diseases, screening for drug metabolism and hepatotoxicity. Thus further investigation of the method to efficiently generate hepatocytes is in great need. Bone Mesenchymal Stem Cells (BMSCs) were collected from rat femurs and tibias. FOXA2 and HNF1α genes were constructed into a lentiviral vector and introduced into BMSCs by a lentivirus-mediated overexpression system. Three weeks after the induction, the expressions of FOXA2 and HNF1α, and liver specific genes were analyzed, and hepatocyte-function related assays were performed. Overexpression of both FOXA2 and HNF1α induced the BMSCs to differentiate into hepatocyte-like cells (HLCs). Hepatocyte-specific gene and protein were detected by RT-PCR, Western Blot and Immunofluorescence. These HLCs also exerted some typical hepatocyte functions such as glycogen storage, indocyanine green absorption and lipid accumulation. The combination of FOXA2 and HNF1α can effectively induce BMSCs to differentiate into HLCs. This is a novel and efficient method to prepare HLCs within a short timeline. © 2016 Springer Science+Business Media Dordrecht

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