Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells

• Дата: 2014
• Автори: Han, G., Wei, W., Zhang, X., Lai, Zh., Chen, Ch.
• Формат: Стаття
• Мова: English
• Опубліковано: Інститут фізіології ім. О.О. Богомольця НАН України 2014
• Назва видання: Нейрофизиология
• Онлайн доступ: http://dspace.nbuv.gov.ua/handle/123456789/148307
• Назва журналу: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Цитувати: Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells / G. Han, W. Wei, X. Zhang, Zh. Lai, Ch. Chen // Нейрофизиология. — 2014. — Т. 46, № 4. — С. 375-380. — Бібліогр.: 11 назв. — англ.

We examined what effects are exerted by expression of the bcl-2 gene and by treatment with nerve growth factor (NGF) on the intensity of apoptosis in cultured pheochromocytoma cells (PC12 cells). Half of these cells were transduced with the bcl-2 gene using lentiviral plasmids, and the respective two groups were denoted as bcl-2-PC12 and control (c) PC12. Then the cells were incubated in a serum-free medium in six different modes. One group of c-PC12 cells was incubated in this medium with no additional agents added, another group was incubated with 1.0 mM H₂O₂ , and the third group was incubated with both 1 mM H₂O₂ and 20 ng/ml NGF (groups 1-3). Cells of the another triad were incubated under the same conditions, respectively, but these were bcl-2-PC12 cells (groups 4-6). The apoptosis rate in each group after 1-h-long incubation was measured using a flow cytometry method. A bicinchoninic acid (BCA) technique was used for estimation of expression of Bcl-2 protein in the cultures. As was observed, the action of H₂O₂ significantly increased the apoptosis rate in both c-PC12 and bcl-2-PC12 samplings, while simultaneous action of NGF considerably attenuated such increases. At the same time, values of the apoptosis rate for bcl-2-PC12 cells were much smaller than the respective values for c-PC12 cells under all the three modes of incubation. In H₂O₂ -treated cultures, the amount of Bcl-2 protein dropped, while the treatment with NGF counteracted such shifts. The content of this protein in the bcl-2-PC12 groups was much higher than in the c-PC12 groups. Thus, transduction with the bcl-2 gene significantly inhibits apoptosis in cultured PC12 cells, and a combined influence of expression of this gene and treatment with NGF produces a synergistic effect.