摘要
目的筛选影响卵巢癌顺铂耐药性的靶点基因。方法从GEO数据库中下载对顺铂敏感和产生抗药性的人类卵巢癌细胞基因表达谱和甲基化谱数据(GSE15709),利用R的相关工具包筛选A2780和A2780/DDP(顺铂耐药卵巢癌细胞系)两类卵巢癌细胞之间的差异表达和差异甲基化的基因;使用DAVID数据库对差异表达基因进行功能富集分析;对同时发生了差异甲基化、差异表达且甲基化水平、表达水平的变化趋势相反的基因,进一步利用q RT-PCR技术检测这些基因在两种卵巢癌细胞中的表达值。结果研究发现在两种卵巢癌细胞之间发生了416个差异表达和281个差异甲基化的基因,这些差异表达基因主要富集于细胞周期、核分裂和蛋白修饰负调控等生物过程。此外,细胞周期、DNA复制和p53等通路在这些基因中同样富集。共发现4个发生了差异甲基化、差异表达且甲基化变化水平、表达水平变化趋势相反的基因,q RT-PCR实验验证了这些基因在两种卵巢癌细胞中的表达水平。结论利用生物信息学和分子生物学相结合的方法,可以筛选出部分影响卵巢癌对顺铂抗药性的基因,为进一步揭示其中的分子机制提供实验参考。
Objective To screen the target genes that contribute to cisplatin resistance in ovarian cancer treatment. Methods Gene expression and methylation profiles of ovarian cancer cells that were sensitive or resistant to cisplatin with accession number GSE15709 were downloaded from GEO database. Differential expressed and methylated genes were identi-fied through associating packages in R. DAVID database to screen the enriched GO terms and pathways of the different ex-pressed genes between A2780 and A2780/DDP. Gene Set Enrichment Analysis （GSEA） of different gene was performed against DAVID database. Genes that exhibited difference in both expression and methylation profiles between the two types of ovarian cancer cells as well as genes that present contradictory profile between expression and methylation were verified via qRT-PCR. Results We found 416 different expressed genes and 281 methylated genes between the two types of ovari-an cancer cells respectively. These differential genes were rich in pathways of cell cycle, DNA replication, nucleus division , p53 signaling , and negative regulation of protein modification process etc. Four genes demonstrated contradictory profile be-tween expression and methylation in the two types of ovarian cancer cells and were verified by qRT-PCR. Conclusion Combination of bioinformatics and molecular biology is useful in the identification of target genes that contribute to resis-tance of cisplatin in ovarian cancer treatment and further reveal molecular mechanism behind it.
出处
《天津医药》
CAS
2015年第10期1108-1111,共4页
Tianjin Medical Journal
作者简介
张玉琪（1971），女，主管药师，本科，主要从事临床药学方面研究
