The Study Of The Hepatocarcinogenesis-related Proteins Of The Mouse Induced By Aflatoxin

In order to further understanding the effect of proteins induced by aflatoxin B1 in the mouse liver, in previous study, two-dimensional electrophoresis and mass spectrometry analysis methods were used to analyze different concentrations of Aflatoxin B1 on the impact of proteins in the mouse liver. The result showed that 35 protein spots changed significantly, of which 5 appeared protein and 7 disappeared proteins were analyzed. The results indicated that these 5 proteins maybe cause liver disease or even cancer, and 7 proteins were involved with the material and energy metabolism in the mouse live, thus all these proteins have a certain effect on the mouse liver. That is to say, Aflatoxin B1 might interact with these 12 proteins and finally affect the mouse liver, and ultimately lead to lesion or even cancer.In order to excluding the err of the two-dimensional electrophoresis, we explored the mRNA variation of atp5b, idh3αand prxⅡby fluorescence quantitative PCR after infection. As we expected, the results were according with the protein analysis. Followingly, we inserted these three genes into the protokaryon expression vetor pET28a, and transformed them into the the host E.coli BL21 (DE 3), resepectively. After induced by 100 mmol/L IPTG, the positive bacteria successfully expressed these proteins. Finnally, the atp5b and reporter gene gfp were inserted into the mammal cell expression vector pCDNA3.1hisB. These work were helpful for revealing the function of atp5b, idh3αand prxⅡin hepatocarcinogenesis induced by Aflatoxin B1.