3-Methylcholanthrene-mediated induction of cytochrome P4501A2 in human hepatoma HepG2 cells as quantified by the reverse transcription-polymerase chain reaction.
3-Methylcholanthrene-mediated induction of cytochrome P4501A2 in human hepatoma HepG2 cells as quantified by the reverse transcription-polymerase chain reaction.
复制标题
通过逆转录聚合酶链反应定量 3-甲基胆蒽介导的人肝癌 HepG2 细胞中细胞色素 P4501A2 的诱导。
DOI：
10.1006/abbi.1994.1413
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发表时间：
1994
影响因子：
3.9
通讯作者：
Bresnick,E
中科院分区：
文献类型：
--
作者：
Chung,I;Bresnick,E
探讨了用HepG 2人肝癌细胞研究细胞色素P4501 A2基因（CYP 1A 2）表达调控的可行性。逆转录-聚合酶链反应（RT-PCR）分析显示，HepG 2细胞组成型表达CYP 1A 2，并能够通过诱导CYP 1A 2 mRNA对3-甲基胆蒽（3 MC）产生应答。在这些研究中，将内含子-外显子连接处的选定序列用作CYP 1A 2和β-肌动蛋白基因的mRNA特异性引物。基于扩增指数期内的两个参数定量诱导水平：产生相同扩增水平的循环数差异和PCR反应效率。使用该方法，估计细胞暴露于3 MC后24 h，CYP 1A 2稳态mRNA水平增加至最大值12倍。在24和48 h时，基础表达的降低以及CYP 1A 2 mRNA积累的增加似乎是总体诱导的原因。这些结果表明，HepG 2细胞系是研究CYP 1A 2表达调控的合适细胞。
The feasibility of using HepG2 human hepatoma cells to study the regulation of the expression of the cytochrome P4501A2 gene (CYP1A2) was examined. The reverse transcription-polymerase chain reaction (RT-PCR) assay revealed that HepG2 cells constitutively express CYP1A2 and are able to respond to 3-methylcholanthrene (3MC) by an induction of CYP1A2 mRNA. In these studies, selected sequences from intron-exon junctions were used as mRNA-specific primers for both CYP1A2 and the β-actin gene. The level of induction was quantitated based on two parameters within the exponential phase of the amplification: the difference in the number of cycles that yields the same level of amplification and the efficiency of the PCR reaction. Using this method, it was estimated that the CYP1A2 steady-state mRNA level increased to a maximum of 12-fold at 24 h after exposure of the cells to 3MC. Both a reduction in basal expression as well as an increased accumulation of CYP1A2 mRNA appeared responsible for the overall induction at 24 and 48 h. These results suggested that the HepG2 cell line would be appropriate for studying the regulation ofCYP1A2expression.