胰岛素和表皮生长因子刺激引起的蛋白激酶B磷酸化与PTEN突变的关系  

Relationship between PTEN mutations and protein kinase B phosphorylation caused by insulin or recombinant human epidermal growth factor stimulation

作　　者：钟海兰[1] 胡先福 林建华[3] 

机构地区：[1]南方医科大学南方医院消化内科,广东广州510515 [2]龙华新区人民医院消化内科,广东深圳518109 [3]南方医科大学南方医院肝胆外科,广东广州510515

出　　处：《细胞与分子免疫学杂志》2016年第8期1065-1068,1072,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81172585)

摘　　要：目的研究第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)突变对人鼻咽癌细胞系CNE-1中蛋白激酶B(Akt)的磷酸化的影响。方法 CNE-1细胞用含100 m L/L胎牛血清RPMI1640培养基培养,并分别转染野生型PTEN(wt PTEN)质粒、突变型PTEN C124S质粒和突变型PTEN G129E质粒。各组细胞加刺激前用无血清RPMI1640培养基饥饿过夜,用0.15 IU/m L胰岛素或0.3μg/m L重组人表皮生长因子(rh EGF)刺激,Western blot法检测Akt的磷酸化水平。结果胰岛素和rh EGF刺激均可导致CNE-1细胞Akt信号激活;wt PTEN可抑制胰岛素或rh EGF刺激引起的Akt信号激活;PTEN C124S突变体可激活胰岛素刺激引起的Akt信号,但不可激活rh EGF刺激引起的Akt信号;PTEN G129E突变体抑制胰岛素刺激引起的Akt信号激活。结论 wt PTEN可抑制胰岛素或rh EGF刺激引起的Akt信号激活,但PTEN C124S和G129E突变体不能一致性激活Akt信号表达。这表明PTEN基因突变可能与Akt信号表达无关。Objective To study the effect of phosphatase and tensin homolog deleted on chromosome 10 （PTEN） mutations on protein kinase B （Akt） phosphorylation of CNE-1 nasopharyngeal carcinoma cell line. Methods CNE-1 cells were cultured in RPM11640 medium containing 100 mL/L fetal calf serum, and then transfected with wild-type PTEN （wtPTEN）, mutant PTEN C124S and mutant PTEN G129E plasmid separately. After overnight serum starvation, the cells were stimulated with 0.15 IU/mL insulin or 0.3μg/mL recombinant human epidermal growth factor （rhEGF）. At last, Akt phosphorylation was evaluated by Western blotting. Results Insulin or rhEGF stimulation led to Akt activation in CNE-1 cells. The wtPTEN inhibited insulin- or rhEGF-stimulated phosphorylation of Akt. PTEN C124S mutant activated insulin-stimulated phosphorylation of Akt, but not rhEGF-stimulated phosphorylation of Akt. PTEN G129E mutant inhibited insulin-stimulated phosphorylation of Akt. Conclusion The wtPTEN inhibited insulin- or rhEGF-stimulated phosphorylation of Akt, while PTEN C124S and G129E mutants failed to activate the phosphorylation of Akt consistently. This suggested PTEN mutations might not be correlated with activated Akt.

关 键 词：第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN) 基因突变 蛋白激酶B(Akt)磷酸化 胰岛素 重组人表皮生长因子(rhEGF) CNE-1人鼻咽癌细胞系 

分 类 号：Q279[生物学—细胞生物学]