【摘要】 目的 :研究ras癌基因和nm2 3抑癌基因的高表达对细胞膜磷脂酶D(PLD)活性的影响。方法 :用ras癌基因和nm2 3抑癌基因构建的重组质粒 ,分别转染人肝癌细胞株 772 1,建立了高表达ras癌基因和nm2 3抑癌基因的H ras 772 1和nm2 3 H 772 1细胞株 ,测定 772 1、mock细胞 (空载体转染的 772 1细胞 )、H ras 772 1和nm2 3 H 772 14种细胞中 ,细胞膜PLD酶活性和PLD酶蛋白表达量。结果 :在H ras 772 1细胞中 ,PLD酶活性比 772 1、mock细胞中PLD酶活性显著升高。nm2 3 H 772 1细胞中 ,酶活性却显著降低。但在二种细胞中 ,PLD酶蛋白的量未见显著变化。结论 :ras癌基因和nm2 3抑癌基因的高表达对PLD酶活性的影响 ,可能主要是通过影响PLD酶活性的调节因素来实现的。更多还原

【Abstract】 Objective:To explore the effects of the high expressions of ras oncogen and nm23 suppressor gene on the activity and protein level of the plasma membrane phospholipase D. Methods:nm23-H/pcDNA3 and H-ras/pcDNA3 plasmids were constructed with pcDNA3 plasmid and nm23-H cDNA or H-ras cDNA respectively. Then the recombinant plasmids were transfected into 7721 human hepatocarcinoma cell line so the nm23-H/7721? H-ras/7721 and mock cell line (pcDNA3 was transfected into 7721 cell) were obtained. PLD activities and PLD protein levels were detected. Results:The high expression p21 ras protein or nm23 protein was shown in H-ras/7721 or nm23-H/7721 respectively with western blot. The PLD activity in H-ras/7721 was much higher than in mock and 7721 (P<0.001).In cantrast,the PLD activity in nm23-H/7721 was significantly lower than in mock and 7721(P<0.001).The PLD protein levels, however, were not significantly different in H-ras/7721,nm23-H/7721,mock and 7721 cell lines. Conclusion:The effects of the high expression of ras oncogene and nm23 suppressor gene on the plasma membrane PLD were probably caused by regulation of PLD activity rather than the expression level of PLD.更多还原