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为了研究PKC在NIH3T3细胞中G2/M期作用,检测了PKC的激动剂1-oleoty-2-acetyl-sn-glycerol（OAG）对G2/M期关键的调控因子MPF活性变化及细胞周期进程的影响。细胞同步化处理用Aphidicolin。用 PI（Propidium Iodide）染料与DNA结合,流式细胞仪检测细胞周期的改变。同时测定OAG加入后G2期 PKC和MPF的活性改变。结果表明Aphidicolin同步在G1/S期的细胞,释放后在经过1-2h的恢复期后,约在7h左右到达早G2期,并在10h达到G2末期。OAG在浓度为50μmol/L,100μmol/L,150μmol/L时都可激活 PKC,而CalphostinC在300nmol/L时最大限度地抑制PKC活性。50μmol/L的OAG至少1150see没有改变细胞内钙。50μmol/L的OAG在G2期使MPF活性降低,而300nmol/L calphostin C可以使MPF活性升高。在G2早期加入 OAG50μmol/L可以引起NIH3T3细胞阻滞在G2/M期交界。
In order to investigate the role of PKC in G2 / M phase in NIH3T3 cells, the effect of PKC agonist 1-oleoty-2-acetyl-sn-glycerol (OAG) on the key regulatory factor MPF activity and cell cycle progression influences. Aphidicolin for cell synchronization. PI (Propidium Iodide) dye and DNA binding, flow cytometry to detect changes in the cell cycle. At the same time, the activity changes of PKC and MPF in G2 phase were determined after adding OAG. The results showed that Aphidicolin synchronized cells in the G1 / S phase, after the release of 1-2h after the recovery period, about 7h to reach the early G2, and 10h to reach the end of G2. OAG activated PKC at concentrations of 50μmol / L, 100μmol / L and 150μmol / L, while Calphostin C inhibited PKC activity at 300nmol / L. 50 micromol / L of OAG at least 1150 do not change intracellular calcium. 50μmol / L OAG decreased MPF activity in G2 phase, while 300nmol / L calphostin C increased MPF activity. The addition of OAG50μmol / L in the early G2 phase can cause NIH3T3 cells to arrest at the G2 / M junction.