Expressions of apoptosis-related proteins in hepatocyte growth factor-transfected bone marrow mesenchymal stem cells under hypoxia conditions

doi:10.3969/j.issn.2095-4344.2015.36.002

Abstract

Abstract:

BACKGROUND: Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cell transplantation, but the mechanism is still unclear.

OBJECTIVE: To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions.

METHODS: (1) Bone marrow mesenchymal stem cells were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem
cells was tested by x-gal staining. (2) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cell scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions for 6 hours.

RESULTS AND CONCLUSION: (1) Transfection efficiency of bone marrow mesenchymal stem cells was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradually increased with hypoxia time (P < 0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P < 0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P < 0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cells was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P < 0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cells can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cells under hypoxia and serum-free conditions.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Hepatocyte Growth Factor, Cell Hypoxia

CLC Number:

R394.2

Cai Wen-qin, Wang Jun-sheng, Su Jin-zi, Jiang Jin-feng, Yao Yue-xian. Expressions of apoptosis-related proteins in hepatocyte growth factor-transfected bone marrow mesenchymal stem cells under hypoxia conditions [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(36): 5746-5752.

Figures/Tables 5

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