胃癌患者抑郁相关基因的基因芯片研究和通路分析

Detection of differently expressed genes of depressive gastric cancer patients by cDNA microarray and pathway analysis

目的应用cDNA芯片技术进行胃癌患者抑郁症状相关基因表达谱差异分析及信号转导通路分析,探讨其可能的作用机制。方法采用Zung抑郁量表判定胃癌患者的精神状态,分为两组,抑郁组10例患者,非抑郁组10例患者。手术切除原发灶肿瘤,30min内液氮保存。Trizol一步法提取细胞总RNA并纯化mRNA,逆转录法分别标记荧光素Cy3和Cy5,合成cDNA探针与人类全基因组芯片杂交,扫描荧光信号图像,分析比较两组差异表达基因,行GeneOntology分类和通路分析。结果基因芯片筛选出1088个基因表达水平发生改变且有统计学意义,包括787个上调基因和301个下调基因。GeneOntology分类提示显著变化的基因网络是"Notch信号传导通路"及"神经肌肉突触传递功能"。BioCarta通路分析得到通路网络中相关基因的表达改变情况,Notch通路中5个基因表达上调,为NOV、CNTN1、YY1、Maml2和Spen,5个基因表达显著下调,为PSEN2、APH1A、PSENEN、TP63和TLE1,神经肌肉突触传递相关基因中2个显著下调,为DTNA和KIF1B。结论肿瘤相关性抑郁可能与能量代谢、细胞骨架、营养因子以及酶等多层次的基因表达改变有关,其中Notch信号传导通路和神经肌肉突触传递功能可能发挥了重要作用。

Objective To investigate the depression-related genes and pathways in gastric cancer patients and discuss its possible mechanism.Methods We used Zung self-rating depression scale（SDS）to determine the mental state of patients initially diagnosed with gastric cancer.Patients were divided into two groups,one group with 10 depressive patients,and the other with 10 non-depressive patients.Tumor tissue cut in the surgery was preserved within 30mins with liquid nitrogen.Use Trizol ＂one-step＂ to extract and purify the total RNA in cells,mark fluorescent Cy3 and Cy5 respectively,and synthesize cDNA probes to hybridize the entire human genome chip,scan fluorescence signal image and analyze the differently expressed genes with the Gene Ontology Classification and the pathway analysis.Results The expression level of 1088 genes changed with significance,including 787 up-regulation genes and 301 down-regulation genes.Gene Ontology analysis indicated significant pathways were Notch signaling pathway and neuromuscular function of synaptic transmission.BioCarta pathway analysis showed the related genes changed in the gene network,up-regulation genes in Notch signaling pathway were NOV,CNTN1,YY1,Maml2,Spen,and down-regulation genes were PSEN2,APH1A,PSENEN,TP63,TLE1.DTNA and KIF1B were down regulated in neuromuscular function of synaptic transmission.Conclusion Cancer-related depression may be associated with multi-levels changes of energy metabolism,the cytoskeleton,nutritional factors,enzymes.Above all,Notch signaling pathway and neuromuscular function of synaptic transmission possibly play an important role.