用基因芯片技术研究胰腺癌肝转移的相关基因群及基功能

Pancreatic cancer has a dismal prognosis due to high incidence of liver metastases after surgical resection. However, there are few investigation works for the molecular mechanism of this phenomenon up to the present. Our investigation focused on screening pancreatic adenocarcinoma relevant gene clusters in the process of oncogenesis, development and liver metastasis using newly built DNA array and suppression subtractive hybridization (SSH) technology. 9 histologically defined head pancreatic adenocarcinoma specimens associated with clinical data were screened for molecular profiling analyses using cDNA microarray that represents a set of 4,096 human genes. Using this methodology, 184 differentially expressed genes were screened out after 9 couples of hybridizations, which including 11 novel human genes. We compare the expression profiles with hierarchical clusterings algorithms and focus on the difference between the two groups of the patients, which reflects the different liver metastasis incidence and prognosis. By using hierarchical clustering analysis, we detected a group of genes which related to patients's clinical outcome,and mainly focusing on peptic hormone related genes, signal-transduction-related genes and cellular skeleton genes. we hope these genes may become markers for pancreatic cancer's clinical prognosis. the nude mouse model of human pancreatic cancer orthotopic transplantation and liver meatasis was established by injecting human pancreatic cancer cell SW1990 . Total RNA were extracted from orthotopic transplantation tumor cells and liver metastasis tumor cells respectively. Then the selection of poly(A) RNA from totol RNA was performed. We performed SSH and constructed the subtractive cDNA library, then screened the library with Dot blot. DNA sequencing and bioinformatics analysis were performed, and 58 metastasis relevant genes were screened out , among which there ard 9 novel genes. We entitled a novel gene S100P, and has obtained the gene bank accession (accession No. AF539739).With thorough functional investigation of gene cluster involving in the process of oncogenesis, development and liver metastasis, which will help us understanding the mechanism of metastasis in pancreatic cancer, and investigating the therapeutic method for pancreatic cancer related recurrence and metastasis.

应用双向抑制消减杂交筛选人胰腺癌及肝转移癌mRNA差异表达基因片段，建立消减cDNA文库并制备成cDNA表达谱芯片，与人胰腺及肝转移癌mRNA荧光标记cDNA探针进行芯片杂交，筛查在胰癌肝转移过程中与肿瘤转移特异性相关的基因并进行功能研究，探索这一基因群的表锲妆浠胫琢鲎频哪谠诹涤牍媛桑芯苛俅苍し酪劝┦鹾笞频闹瘟品椒ㄌ峁├砺壑傅肌?

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