不同周期Sf9细胞琥珀酸脱氢酶酶活的研究

Succinic dehydrogenase activity of Sf9 cells in different cell cycle phases

为了研究不同周期Sf9细胞线粒体琥珀酸脱氢酶酶活的差异,首先使用TdR(胸腺嘧啶核苷)双阻断法将Sf9细胞同步化于G1/S期,恢复生长分别得到S、G2/M、G1期的细胞,然后利用MTT法检测不同细胞周期的Sf9细胞琥珀酸脱氢酶酶活,发现处于G1期的细胞酶活比S、G2/M期的细胞酶活大.跟踪细胞成长,在不同的时间点采样,以MTT法检测琥珀酸脱氢酶酶活,发现接种后48 h的Sf9细胞酶活最大,此时与其他时刻相比的Sf9细胞处于G1、S期所占的比例最大.

To investigate that the cell cycle phase might play a role in the succinic dehydrogenase activity, we firstly blocked Sf9 cells at G1/S phase boundary. The cells in S, Gland G2/M phase could be obtained after those cells are released. Suceinic dehydrogenase activity of Sf9 cells in different cell cycle phases was tested by MTT assay. We noted that Sf9 cells in G1 phase has strongest suecinic dehydrogenase activity compared to Sf9 cells in S and G2/M phase, stronger in S phase but smallest in G2/M phase. Secondly, to test the suecinic dehydrogenase activity of Sf9 cells by MTT assay as cells growth was proceeding. We noted that the succinic dehydrogenase activity was also biggest when Sf9 cells entered into the exponential growth phase by 24 hour cell, i.e. 48 hour of post inoculation in this experiment. And we also found that the sum of the percentage of cells in G1 and S phase was biggest.