摘要
目的:喹烯酮能诱导HepG2细胞S期阻滞,引起DNA和染色体损伤。本研究旨在对喹烯酮致HepG2细胞S期阻滞机制进行筛选和分析。方法:本研究对喹烯酮染毒后HepG2细胞和未染毒HepG2细胞进行了Agilent全基因组的表达谱芯片检测,应用Genespring软件对差异表达的基因进行相关通路分析,并使用荧光定量PCR技术对部分DNA复制相关表达差异基因进行验证。结果:在芯片点样的41000个寡聚核苷酸片段中,喹烯酮染毒HepG2后,差异表达基因共1750个,其中表达上调的基因共446个,表达下调的基因共1304个。差异表达基因通路分析结果表明,与细胞周期、MAPK通路、DNA复制及DNA修复等通路相关的基因表达差异显著。经荧光定量PCR验证,DNA复制相关基因表达变化趋势与芯片检测结果相一致。结论:DNA复制相关基因的下降导致S期DNA复制的不完全性以及DNA损伤后引起的DNA修复,使得HepG2细胞在喹烯酮染毒后引起S期阻滞。基因表达谱芯片检测分析的结果为喹烯酮致HepG2细胞S期阻滞机制的深入研究奠定了基础。
OBJECTIVE：Quinocetone induced S phase arrest,caused DNA and chromosome damage in HepG2 cells.Our purpose was to screen and analyse the mechanism of S phase arrest induced by Quinocetone. METHODS：The Agilent whole genome gene expression microarray was used to detect gene expression profile of HepG2 cells treated and untreated with Quinocetone.Futhermore,pathway analysis by Genespring software was carried out based on the differentially expressed genes,real time qPCR was used to verify some dramatically changed genes in DNA replication process.RESULTS：1 750 differentially expressed genes,with 446 genes up-regulated and 1 304 genes down-regulated,were found in HepG2 cells treated with Quinocetone in the total of 41 000 oligonucleotide fragments. Pathway analysis revealed that the differentially expressed genes gathered in functional pathways like cell cycle checkpoint,MAPK signaling pathway,DNA replication and DNA repair,etc.Moreover,the results of real time qPCR focused on the changed DNA replication genes were consistent with cDNA microarray.CONCLUSION：The down-regulation of DNA replication genes caused incomplete DNA synthesis,and the DNA repair after its damage.These two events lead to S phase arrest after Quinocetone treatment.Moreover,the whole genome cDNA microarray analysis provided us much useful information for further research on the mechanism of S phase arrest induced by Quinocetone.
出处
《癌变·畸变·突变》
CAS
CSCD
2011年第5期325-329,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
教育部博士点新教师基金(20100008120003)
