摘要
利用化疗药物5-氟尿嘧啶(5-Fu)对胃癌细胞株进行筛选和诱导,建立具有耐药性的胃癌细胞株.与亲代细胞株对比生长特性及基因表达谱,初步探讨胃癌细胞的耐药机制.采用四唑盐比色法(MTT)测定5-Fu对胃癌细胞株BGC-823的半数抑制浓度(IC50);根据IC50设计5-Fu剂量,用大剂量的5-Fu逐渐递增间歇给药的方法,反复筛选,获得5株胃癌耐药细胞株.比较耐药细胞株和亲本细胞株的形态和生长特性,继而再用人类肿瘤基因芯片(human cancer arrays)比较测试亲本与耐药细胞株的基因表达谱差异;对其中189个上调基因和133个下调基因采用Gene Ontology中的BP(Biological Process)分析这些基因相关的功能,并用MAS2.0分析这些基因可能涉及的信号通路.结果提示,胃癌细胞株耐药(5-Fu)相关基因可能与转录因子信号转导、TNF受体介导的细胞凋亡和抗凋亡、细胞周期调节、细胞粘附及MAP激酶类的功能相关.对细胞周期、信号转导相关的6个基因采用定量PCR验证,结果与基因表达芯片结果一致.上述结果有利于进一步发现胃癌细胞的耐药基因和相关信号通路,探索抗胃癌治疗的耐药机制.
The resistant gastric cancer cell lines were established by exposing cells to chemotherapeutical drug 5-flurouracil（5-Fu） with repeated inducing and screening.We compared the resistant cell lines to their parent cell line Bgc-823 on cell growth and gene expression profile.The comparisons might providesome indications to explore the mechanisms of drug resistance to gastric cancer cell line.The MTT assay wasused to detect 50 percent inhibiting concentration（IC50） of BGC-823.The concentration of 5-Fu wasdetermined in the culture dishes of BGC-823 according to the value of the IC50.The resistant cell lines（5strains） were obtained by adding high concentration 5-Fu,and increasing the drug concentration withinterval,repeatedly and gradually.We compared the features of cell growth and expression profiles between the resistant cells and BGC-823（parent cell line） using ＂Human Cancer Arrays＂.The functional BP （Biological Process）analysis of Gene Ontology was used to classify 189 up-regulated genes and 133 down-regulated genes.These differential expressional genes were also analyzed by MAS2.0 tool to show genes andtheir relationships in signaling pathways.The results indicated that the resistant genes might be associatedwith transcription factors and their signaling transductions,apoptosis and anti-apoptosis by TNF receptors,regulations of cell cycles,cell adhesion and MAP kinase.Six genes related to cell cycle and signaltransductions were selected to testify expression microarrays with RT-qPCR.The results of RT-qPCRcoincided with those of microarrays.The established resistant gastric cell lines would be beneficial forfurther finding the resistant genes and relative signaling pathways,and mechanisms of drug resistance.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2011年第2期180-189,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
内蒙古医学院博士课题启动资金资助项目~~
关键词
5-氟尿嘧啶
耐药性
胃癌细胞株
基因表达谱
5-flurouracil
drug resistance
cell lines of gastric carcinoma
gene expression profiles
