摘要
背景与目的:由己建立的小鼠基因打靶模型证实,K-ras突变启动了胰腺癌前病变即胰腺导管上皮内瘤变(pancreatic intraepithelial neoplasia,PanIN),Smad4基因失活可促使PanIN细胞恶性转化。PanIN细胞中Smad4基因静默后其下游基因表达将如何改变及促使PanIN细胞恶性转化,目前尚不清楚。基于此目的,我们在己成功分离建立K-ras突变启动的PanIN细胞,并应用siRNA干扰技术静默PanIN细胞株中内源性Smad4表达的基础上,应用全基因表达谱芯片研究Smad4基因静默前PanIN细胞与其静默后PanIN-S细胞基因表达谱的变化。方法:应用包含41 174个小鼠基因转录本Agilent小鼠4×44K全基因芯片检测Smad4基因静默前后PanIN细胞基因表达谱变化,分析差异表达基因,并选择其中有差异表达的部分基因进行实时荧光定量多聚酶链反应(Real-time PCR)验证。结果:基因芯片筛选出差异倍数2倍以上的差异表达基因,如Fut9、CXCR4、MMP2、PIK3C2G、Cplx4、Lemd1、TIMP4、Reln和PITX2等。Real-time PCR验证差异表达基因CXCR4、MMP2、PIK3C2G、TIMP4、PITX2与基因芯片结果一致。结论:Smad4基因静默前后PanIN细胞中CXCR4、MMP2、PIK3C2G、TIMP4、PITX2基因表达存在显著差异,其中CXCR4、PIK3C2G基因可能与胰腺肿瘤新生血管形成相关,MMP2、TIMP4基因可能与胰腺肿瘤细胞黏附、浸润、迁移相关,PITX2表达高低可能与胰腺肿瘤预后相关。
Background and purpose： The mutation of K-rasGI2D bring about pancreatic intraepithelial neoplasia （PanIN）, which is thought to be a precursor lesion of pancreatic ductal adenocarcinoma via the research in gene targeted mice model. PanIN with the silencing of Smad4 could progress to invasive adenocarcinomas. The change about the genes expression profile between PanIN cells and PanIN cells with Smad4 silencing （PanIN-S cells） which promoted PanIN cells malignant transformation still remains poorly understood. In our previous research, PanIN-S cells were established by stable transfection with leutiviral mediated Smad4 RNA interference. In the present study, we aimed to explore the gene differentially expressed genes between PanlN ceils and PanIN-S cells. Methods： A gene microarray （Agilent） with 41 174 transcripts was employed to examine the gene differential expression in PanlN cells and PanIN-S cells. The differentially expressed genes were identified and subjected to Real-time PCR analysis. Results： A large number of differentially expressed genes whose fold change was larger than 2 were examined, embracing Fut9, CXCR4, MMP2, PIK3C2G, Cplx4, Lemdl, TIMP4, Rein, PITX2 and so on. The fold change of Real-time PCR of CXCR4, MMP2, PIK3C2G, TIMP4, PITX2 was consistent with that of gene expression profiling. Conclusion： The outcome of the gene expression profiling indicated many differentially expressed genes associated with PanlN cells and PanIN-S cells. CXCR4, PIK3C2G may be related to the pancreatic tumorigeness via angiogenesis. MMP2, TIMP4 may be bound up with adhesion, invasion and migration of cancer cells. The expression levels of PITX2 may be associated with prognosis of pancreatic cancer.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2011年第1期1-5,共5页
China Oncology
基金
国家自然科学基金资助项目(No:30672385)
国家自然科学基金资助项目(No:30971130)
