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目的:利用基因芯片技术,对体外筛选获得的耐不同浓度5-氟尿嘧啶(5-FU)的胰腺癌细胞株基因表型进行对比,找出不同耐药表型细胞株的基因表达改变情况,以此发现一些与胰腺癌细胞5-FU耐药相关的重要基因或通路。方法:用不断增加浓度的5-FU反复刺激细胞,直到细胞能完全耐受不同浓度5-FU,获得耐不同浓度5-FU的MIAPaCa-2细胞株。由基因芯片筛选出明显改变的基因后再用实时定量PCR方法验证。结果:本研究成功地获得两种对5-FU耐药的胰腺癌细胞株,低耐药表型MIA-FU-2.4和高耐药表型MIA-FU-10.0。基因分析结果提示耐药可能与广泛的基因表达改变相关。在MIA-FU-2.4和MIA-FU-10.0两种表型中分别有1075及1628个基因出现了表达改变,涉及细胞周期、细胞黏附、信号转导、DNA修复和凋亡等多种生物学功能。结论:胰腺癌MIAPaCa-2细胞株5-FU耐药的发生涉及多种细胞生物学功能改变,特别与凋亡、DNA修复、细胞周期等基因和一些信号转导途径相关。
OBJECTIVE: To compare the gene phenotypes of pancreatic cancer cell lines with different concentrations of 5-fluorouracil (5-FU) screened by in vitro screening to find out the gene expression changes of different resistant phenotype cell lines This found that some of the pancreatic cancer cell 5-FU resistance-related genes or pathways. METHODS: The cells were repeatedly stimulated with increasing concentrations of 5-FU until the cells were completely tolerated with different concentrations of 5-FU to obtain MIAPaCa-2 cell lines with different concentrations of 5-FU. By gene chip screening of significantly altered genes and then real-time quantitative PCR method validation. Results: Two pancreatic cancer cell lines resistant to 5-FU, MIA-FU-2.4 with low resistance and MIA-FU-10.0 with high resistance were successfully obtained in this study. Genetic analysis suggests that resistance may be related to a wide range of gene expression changes. In the two phenotypes of MIA-FU-2.4 and MIA-FU-10.0, 1075 and 1628 genes appeared to be altered in expression, involving various biological processes such as cell cycle, cell adhesion, signal transduction, DNA repair and apoptosis Features. CONCLUSIONS: The multidrug resistance of MIAPaCa-2 cells to pancreatic cancer involves a variety of cellular biological functions, especially related to genes such as apoptosis, DNA repair, cell cycle and some signal transduction pathways.