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Archive > Volume 22 Issue 4 > 2015,22(4):448-453. DOI:10.3872/j.issn.1007-385X.2015.04.007 Prev Next
Basic Research
Enforced expression of Bestrophin 3 promotes apoptosis of hepatocellular carcinoma cell HepG2
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Abstract:
Objective:To investigate the effect and mechanism of Bestrophin 3 on the apoptosis of human hepatocellular carcinoma cell HepG2. Methods: HepG2 cells were transfected with various multiplicity of infection (MOI=10, 20, 40 and 80) of adenovirus expressing Bestrophin 3, LacZ, or control. The infection efficiency of the adenovirus was measured by Western blotting. Proliferation and apoptosis rates of the transfected cells were measured with CCK-8 assay and flow cytometry. Effects of Bestrophin 3 on the expression of Bcl-2 and Bax and the release of cytochrome C (Cyt C) were determined by using immunoblotting. Influence of Bestrophin 3 overexpression on mitochondrial membrane potential was assessed by JC-1 and fluorescence microscopy. Results: The expression of Bestrophin 3 reached the maximal level after adenoviral infection at 40 MOI. Compared with the control and LacZ-expressing groups, the cell viability of the Bestrophin 3-expressing group was significantly reduced \[(79.37±1.76)% vs (98.67±3.02)% and (99.67±3.25)%，P<005\], whereas the apoptosis rate was significantly increased \[(29.47±2.37)% vs （5.47±0.37）% and (4.95±044）%，P<0.05\]. In the cells infected with the adenovirus expressing Bestrophin 3, there were decreased level of Bcl-2 and increased level of Bax, leading to a significant reduction of Bcl-2/Bax ratio. In HepG2 cells overexpressing Bestrophin 3, the mitochondrial membrane potential was also significantly lower compared with that of controls (1.00±0.00)% vs 0.64±0.09%, P<0.05), and Cyt C content was reduced in mitochondria and elevated in cytoplasm (all P<0.05). Conclusion: Enforced expression of Bestrophin 3 increases the apoptosis of HepG2 through reducing Bcl-2/Bax ratio and promoting cytochrome C release from the mitochondria.
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Project Supported:
Project supported by the Natural Science Foundation from the Education Bureau of Henan Province（No. 13A320854）
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