Affymetrix基因表达谱芯片技术筛选胰腺癌异常表达基因的研究

Screening of abnormally expressed genes in pancreatic cancer by Affymetrix gene expression chip

目的:利用基因表达谱芯片筛选出胰腺癌组织与癌旁正常组织的差异表达基因。方法:收集新疆医科大学第三附属医院2014年1月—2016年6月间手术切除的10例胰腺导管细胞癌组织及其相应癌旁正常组织,用TRIzol法提取总RNA后采用含49395个基因探针的Affymetrix基因表达谱芯片筛选出差异表达基因,并行GO分析和pathway分析,对部分差异表达基因行实时定量PCR法验证表达情况。结果:样本总RNA检测合格,基因芯片质量评估良好,检测结果可靠且可重复性高。经芯片降噪处理后共检测38079个基因,其中存在差异表达的基因共512个,表达上调的基因419个,表达下调的基因93个;共287个差异表达基因编码与3个GO分类(生物学过程、分子功能、细胞组分)相关的蛋白;共29条信号通路存在明显基因差异表达,涉及126个基因。经PCR验证,差异表达基因CPB1、CELA3B、CPA1、POSTN、PLA2G1B、CTRC及SPINK1的表达情况与基因芯片检测结果吻合。结论:胰腺导管细胞癌组织与癌旁正常组织间存在大量差异表达的基因,这些基因主要与生物学过程、分子功能及细胞组分相关,且参与了多个细胞信号通路的调控。

Objective： To screen the differentially expressed genes in pancreatic carcinoma versus paired normal adjacent tissue by gene expression profile chip. Methods： The surgical specimens of pancreatic ductal cell carcinoma and adjacent normal pancreatic tissue from 10 patients from January 2014 to June 2016 in the Third Affiliated Hospital of Xinjiang Medical University were collected, and differentially expressed genes between them were screened by an Affymetrix gene expression profile chip containing 49 395 gene probes after total RNA extraction by TRIzol method, and then, GO and pathway analyses were performed. Finally, some differentially expressed genes were verified by real-time PCR. Results： The total RNA of samples passed the test, gene chip assessment yielded high-quality information and results of the test were reliable and highly repeatable; 38 079 genes were examined after denoising process of the chips, involving 512 differentially expressed genes that included 419 up-regulated genes and 93 down- regulated genes; there were 287 differentially expressed genes encoding proteins that were related to the three GO ontologies （biological process, molecular function and cellular component）, and 29 signaling pathways were associated with significant differential gene expressions, involving 126 genes. The expressions of differentially expressed genes CPB1, CELA3B, CPA1, POSTN, PLA2G1B, CTRC and SPINK1 determined by real-time PCR were consistent with the results ofgene chip analysis. Conclusion： qlaere are a large number of differentially expressed genes between pancreatic ductal cell carcinoma and normal adjacent pancreatic tissue. These genes are mainly associated with biological processes, molecular functions and cellular components, and also involved in the regulation of multiple cellular signaling pathways.