Molecular Biological Analysis of Cells and Tissues from Animals Exposed to Chemicals and Ultraviolet Rays

该项目旨在从分子上阐明细胞和组织在暴露于化学物质和紫外线下的反应。我们首先将氯仿作为一种生物有害化学物质，并测定了暴露于氯仿的小鼠各组织中的转录本。采用差异显示法，分别在玉米油培养皿和单独玉米油培养皿中腹腔注射氯仿的ICR小鼠脏器中获得mRNA指纹。比较指纹图谱，证实两个mRNA片段是由肾脏暴露氯仿诱导的。对这些mRNA片段进行测序并鉴定为ADAMTS-1（一种具有血小板反应蛋白基序的崩解素和金属蛋白酶-1）和内源性MuLV（小鼠白血病病毒）mRNA的一部分。然后采用定量RT-PCR方法检测氯仿小鼠肾、肝和脑中ADAMTS-1和MuLV mrna的含量。有趣的是，ADAMTS-1和MuLV mrna仅在肾脏中增加，而在肝脏和大脑中没有增加。这些结果表明，组织对氯仿的反应是组织特异性的。由于ADAMTS-1基因被细胞因子激活，肾脏ADAMTS-1 mRNA的增加可能是通过氯仿激活的细胞因子系统继发的现象。MuLV可能以原病毒的形式存在于基因组中，其在肾脏中的激活为我们提供了与氯仿相关的癌症的新线索。对其他化学物质和紫外线的研究正在进行中。总之，我们确定了暴露于氯仿的小鼠的ADAMTS-1和MuLV基因的肾脏特异性激活。

The project aimed to melecularly clarify the reactions of the cells and tissues when exposed to chemicals and ultraviolet rays. We took up chloroform as a bio-hazardous chemical first and determined the transcripts in various tissues of mice exposed to chloroform. We obtained mRNA fingerprints, by using differential display method, from the organs in ICR mice intraperitoneally injected with chloroform in corn oil vehicle or corn oil vehicle alone.Compared the fingerprints, two mRNA fragments were proved to be induced by chloroform-exposure in kidney. These mRNA fragments were sequenced and identified as the parts of ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs-1) and endogenous MuLV (murine leukemia virus) mRNAs. Then we measured the amounts of ADAMTS-1 and MuLV mRNAs in kidney, liver and brain of mice exposed to chloroform by using quantitative RT-PCR.Interestingly, ADAMTS-1 and MuLV mRNAs increased only in kidney, but not in liver and brain. These results indicated that tissues reacted to chloroform in a tissue-specific manner. As ADAMTS-1 gene is activated by cytokines, the increase in ADAMTS-1 mRNA in kidney may be a secondary phenomenon through the cytokine systems activated by chloroform. Activation of MuLV,which may be present in the genome as provirus, in kidney gives us new clues to chloroform related-cancers. The research of other chemicals and ultraviolet rays are on the way. In conclusion, we identified kidney-specific activation of the ADAMTS-1 and MuLV genes of mice exposed to chloroform.