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目的探讨转染磷脂酰乙醇胺N-甲基转移酶2(PEMT2)基因抑制大鼠肝癌CBRH-7919细胞增殖的机制。方法采用免疫细胞化学和蛋白质印迹法观察pemt2过表达对肝癌细胞不同亚型蛋白激酶C(PKC)表达及在细胞内转位的影响,同时采用高效薄板层析技术对细胞内二脂酰甘油(DAG)的水平进行检测。结果转染PEMT2使细胞CPKC α表达降低,CPKC β2表达增加,并由胞浆向质膜转位。同时细胞内DAG水平下降。转染PEMT2对其它PKC亚型的表达及细胞内转位无显著影响。结论转染PEMT2 对不同亚型PKC表达及细胞内转位的影响可能与其抑制细胞增殖,诱导凋亡的机制有关。
Objective To investigate the mechanism of PEMT2 transfection inhibiting the proliferation of rat hepatocellular carcinoma cell line CBRH-7919. Methods Immunocytochemistry and Western blotting were used to observe the effect of pemt2 overexpression on protein kinase C (PKC) expression and intracellular translocation in hepatocellular carcinoma cells. Meanwhile, DAG) levels were detected. Results Transfection of PEMT2 decreased the expression of CPKCα and increased the expression of CPKC β2, and translocated from the cytoplasm to the plasma membrane. At the same time, intracellular DAG levels decreased. Transfection of PEMT2 had no significant effect on the expression of other PKC isoforms and intracellular translocation. Conclusion The effect of transfection of PEMT2 on PKC expression and intracellular translocation in different subtypes may be related to its mechanism of inhibiting cell proliferation and inducing apoptosis.