摘要
目的基于磷酸化蛋白组学研究,探讨代谢信号通路异常与垂体生长激素(GH)腺瘤的相关性。方法垂体GH腺瘤标本取材于2010年8月至2017年8月于首都医科大学附属北京天坛医院神经外科行手术治疗的12例患者,正常垂体组织取材于6例因交通意外死亡尸体捐献者。采用超高效液相色谱和蛋白质质谱分析垂体GH腺瘤与正常垂体组织间的差异表达蛋白质,进一步通过生物信息分析方法识别与垂体GH腺瘤代谢相关的信号通路;采用免疫蛋白印迹方法验证该信号通路中关键分子在垂体GH腺瘤和正常垂体组织中的表达。结果蛋白质谱分析结果显示,垂体GH腺瘤与正常垂体组织间存在1213个差异磷酸化蛋白位点(P<0.05,差异倍数≥1.5或<0.67)。在垂体GH腺瘤中,表达上调的磷酸化蛋白位点有342个,表达下调的磷酸化蛋白位点有871个。这1213个差异磷酸化蛋白位点分布在667个蛋白中。信号通路分析(IPA)生物信息学结果显示,667个差异表达的磷酸化蛋白质主要富集在腺苷酸活化蛋白激酶(AMPK)信号通路和糖代谢相关信号通路(P<0.05)。在糖代谢通路富集的10个磷酸化蛋白中,有9个在垂体GH腺瘤中低表达。信号通路分析显示,AMPK信号通路活性抑制。免疫蛋白印迹结果证实,在垂体GH腺瘤与正常垂体组织间AMPK蛋白质的表达未见明显差异(相对表达量为1.75±0.36对比1.59±0.75,P=0.663),而在垂体GH腺瘤中磷酸化AMPK(ser496)蛋白质表达明显低于在正常垂体组织中的表达(相对表达量为1.47±0.58对比0.32±0.16,P=0.016)。结论磷酸化蛋白质组学分析显示,垂体GH腺瘤存在异常代谢。AMPK信号通路的低表达可能与垂体GH腺瘤异常糖代谢相关。
Objective To identify the role of abnormal metabolism signaling pathways in growth hormone-secreting pituitary adenomas (GH-PAs) based on phosphoproteome profiles. Methods In this study, twelve GH-PAs specimens were collected from patients who were surgically treated at Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University from August 2010 to August 2017. Normal pituitary tissues were collected from 3 body donors who died from traffic accidents. The differential phosphoprotein expression patterns involved in GH-PAs were investigated by nano LC-MS/MS in a group of samples. The phosphoprotein expression data were analyzed by bioinformatics. The expression levels of the candidate phosphoproteins were validated by western blot analysis in another group of samples. Results A total of 1 213 phosphorylated protein sites corresponding to 667 proteins were significantly different between GH-PAs and healthy pituitary glands (P<0.05, FC≥1.5 or <0.67). Among those phosphorylated sites, 342 exhibited lower levels of phosphorylation in GH-PAs and 871 exhibited lower levels of phosphorylation in GH-PAs. The 1 213 phosphorylated protein sites were mapped to 667 phosphoproteins. The majority of differentially expressed phosphorylated proteins were significantly enriched in glycolysis and the AMPK signaling pathway in GH-PAs by IPA (P<0.05). In glycolysis pathway, 9 of 10 phosphorylated proteins exhibited lower phosphorylation levels in GH-PAs than healthy pituitary glands. The AMPK signaling pathway was demonstrated to be inhibited in GH-PAs by Pathway Activity Analysis. Notably, western blot proved that the level of AMPK(1.75±0.36 vs. 1.59±0.75, P=0.663) had no difference and the phosphorylated levels of AMPK (ser496) was significantly lower in GH-PAs than in healthy pituitary glands(1.47±0.58 vs. 0.32±0.16, P=0.016). Conclusion Phosphorylation proteomics revealed the abnormal metabolism in GH-PAs. Decreased phosphorylation of the AMPK pathway may be critical for glucose metabolism in GH-PAs.
作者
赵斯达
冯洁
李储忠
何乐
张亚卓
Zhao Sida;Feng Jie;Li Chuzhong;He Yue;Zhang Yazhuo(Beijing Neurosurgical Institute,Capital Medical University,Belting 100070,China)
出处
《中华神经外科杂志》
CSCD
北大核心
2019年第1期73-77,共5页
Chinese Journal of Neurosurgery
基金
国家自然科学基金(81502389).
关键词
分泌生长激素的脑垂体腺瘤
蛋白质组学
代谢
信号传导
Growth hormone-secreting pituitary adenoma
Proteomics
Metabolism
Signal transduction
