| Objective To study the effects of B(a)P to the promoter region methylation of p16andRASSF1A genes in primary culture trachea-bronchia epithelial cells of rat in different dosesand at different time. Methods (1) The rat trachea-bronchia epithelial cells were cultured bytissue explant adherent method;(2) MTT reduction assay was used to observe the influence ofa series of B(a)P on proliferation activity of trachea-bronchia epithelial cells of rat;(3) Thetechnique of methylation specific polymerase chain reaction (MSP) was used to analyze themethylations of p16and RASSF1A genes promoter region in the rat trachea-bronchiaepithelial cells by different concentrations of B(a)P. Results (1) This experiment has beensuccessful in cultivating the trachea-bronchia epithelial cells of rat.(2) The results of MTTshowed that the inhibition rate of trachea-bronchia epithelial cells was less than50%on theconcentration of10.0μmol/L or less of B(a)P in the72h, and B(a)P had markedly inhibitedproliferation effects with dose-dependent manner.(3) The primary trachea-bronchialepithelial cells of rat was cultured with B(a)P in the concentration of0.1,1.0,10μmol/L,respectively. There were no methylation of p16and RASSF1A genes promoter region after24h,48h,72h. The results of MSP were tested by sequencing. Conclusion (1) It is a bettermethod to culture the trachea-bronchia epithelial cells of rat with tissue culture method and islow-cost, convenient and practical.(2) Toxicity of B(a)P to rat trachea-bronchia epithelialcells increased accompany with increased of concentration of B(a)P, which had markedlyinhibited proliferation effects with dose-dependent manner.(3) It can not cause methylationof p16and RASSF1A genes promoter region, the primary trachea-bronchial epithelial cells ofrat was cultured with B(a)P in the concentration of0.1,1.0,10μmol/L after24h,48h,72h,respectively. |
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