Influence Of Formaldehyde On Gene Expression Of Rab26 And CDKN2B In Vitro

In recent years, along with the enhancement of living level,household and official decoration has been flourishing too, while people pursuing happy life,troubles caused by the deleterious substance which reside in the decorating materials appeared. there is a little formaldehyde not combined in these materials,it may pollute the indoor air when release from the decorating products and is harmful to human body. Because of its characteristic of extensive sources,serious toxicities,high-level pollution and long-term duration, formaldehyde is considered to be one of the most common pollutants. And the study of formaldehyde has become a lot and difficult spot of environmental and hygienical fields. It is very important to expore the genotoxity of formaldehyde, toxicity mechanism and repair for understanding formaldehyde genotoxicity as well as nosogenesis completely and systematically. In addition, this research will benefit a lot to estimate and improve the indoor air quality by providing scientific foundation and make a safe environmental and occupational concentration standard. This project applied KC1-SDS assay , PCR and RT-PCR,and biological information software of origin and gene-tool,choose HEK293 cell and Hela cell as experimental material to study DPC effect caused by formaldehyde and it's genotoxicies. It mainly includes three parts as follows.1. DPC effect caused by HCHOThe DPC (DNA-protein crosslinks) has a background level in normal cells, there is a low level of DPC in normal cell, this is normal links between DNA and protein or it is metabolic product in organism. When environmental pollutants or chemical careinogens (for example formaldehyde) effect on macro-molecules of lives, excessive DPC will be induced,DPC is formed in the course of duplication and transcription due to it only occurs on single strand of DNA, so DPC will affect the conformation and the function of DNA (duplication and transcription). Also comparing with other damages repairing of DPC is more difficult, Therefore many important genes (eg.tumor suppressor gene) may lost easily during duplication and transcription.In this project we used HEK293 cell as material, treated with HCHO(≥250μM) of different concentration, and studied the DPC effect with SDS-KC1 assay. The result showed that accompany elevating the concentration of HCHO the amount of DPC significantly enhanced(P<0.01),So we took 250umol/L(and higher than 250umol/L) as treating concentration.2. Influence of DPC on duplicationSince 1969 Brutlag and other scientists firstly reported that DPC was produced after chromosome was treated with formaldehyde, most of researches ware concentrate on mechanism of formation and chemical properties,but few reported on how to perform its hereditary toxicities,a paper once reported that genes expressed abnormally in epithelial cells of P344 mouse nasal respiratory tract after treated with formaldehyde ,of which expression of rab26 and CDKN2B gene were down-regulated. In our study we took the rab26 gene in HEK293cell and CDKN2B gene in HeLa cell as the experimental object, The result showed that accompany elevating the concentration of HCHO the amount of DPC significantly enhanced(P<0.01), at the meantime we found that coupled with the concentration of HCHO and the amount of DPC enhanced the amount of replication of Rab26 reduced and the amount of CDKN2B gene changed unsignificantly by PCR3. Influence of DPC on transcriptionOur study took took the rab26 gene in HEK293cell and CDKN2B gene in HeLa cell as the experimental object, treated with HCHO,and applied RT-PCR assay estimated its variation in transcriptional level. The result showed that accompany elevating the concentration of HCHO the amount of DPC significantly enhanced(P<0.01), at the meantime we found that coupled with the concentration of HCHO and the amount of DPC enhanced the amount of replication of Rab26 and CDKN2B gene both reduced...