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作 者:李亚丽[1] 邵彦华 刘之力 夏泉[1] 马克里[1]
机构地区:[1]大连医科大学生物化学教研室,大连116027 [2]大连宝生物公司开发部 [3]大连皮肤病医院
出 处:《中华肝脏病杂志》2006年第7期514-516,共3页Chinese Journal of Hepatology
基 金:国家自然科学基金(39670809)
摘 要: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。方法采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和Western blot方法,观察转染PEMT2基因对大鼠肝癌CBRH 7919细胞磷脂酶C γ 1(PLC γ 1)磷酸化及在细胞内转位的影响;同时观察转染PEMT2基因对肝细胞生长因子受体(c- Met)自身磷酸化活化的影响。结果转染PEMT2后,质膜结合的PLC γ下降,为对照组的45%。膜结合的磷酸化PLC γ 1约为对照组细胞的27%,同时c-Met磷酸化程度显著下降,约为对照组细胞的32%。结论转染PEMT2基因可抑制细胞PLC γ 1磷酸化及由胞浆向质膜转位,抑制c-Met自身磷酸化活化,从而下调CBRH-7919细胞c-Met/PLC γ 1信号转导途经。Objective To explore the mechanism of CBRH-7919 cell proliferation inhibition by transfecting phosphatidylethanolamine N-methyltransferase 2 gene (PEMT2). Methods The effects of PEMT2 transfection on phosphorylation and translocation from cytosol to plasma membrane of PLCγ1 in cells were studied using SDS-PAGE and Western blot techniques. The phosphorylation and activity of c-Met were determined. Results After transfection of pemt2, the PLCγ1 and phosphorylated PLCγ1 conjugated with plasma membrane were decreased by 45% and 27% of that of control cells respectively, and the phosphorylated c-Met was decreased to 32% of that of control cells. Conclusions Transfection of phosphatidylethanolamine Nmethyltransferase 2 gene can inhibit the phosphorylation and translocation from cytosol to plasma membrane of PLCγ1 in cells. At the same time, the autophosphorylation of c-Met was decreased, which suggests that transfection of phosphatidylethanolamine N-methyltransferase 2 gene can downregulate the c-Met/PLCγ1 signaling pathway in CBRH-7919 cells.
关 键 词:癌 肝细胞 大鼠 磷脂酰乙醇胺N-甲基转移酶2 磷脂酶Cγ1
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