The Function Of Isocitrate Dehydrogenase In Aflatoxin Induced Hepatoma

Many researchs found that, the protein level of IDH3α would increase and thelevel of this protein would rised in the hepatoma patients after treatment withaflatoxin B₁. In this experiment, pET28(a)-idh3α recombined plasmid was constructedand the target protein IDH3α was purified frome the expressed products in E. coil.Then polyclone antibody has been obtained from immuned mice and the titer is up to32000.Aflatoxin B₁was extracted from A. flavus via methanol after30d. Theconcentration of aflatoxin B₁was about0.5mg/mL. The extracted aflatoxin B₁0.25mg/mL was used to treated rat by i.p twice a week for4months. During this period,the rats' weight changed in the aflatoxin group were significantly slowly compared toothers group. The bleed of different groups send to hospital for determination of manyliver function indicators. Result showed that ALT, AST, LDH and TBIL in alfatoxingroup were significantly increased than other two groups but DBIL and TP did notchange. The tissue section of livers was been made from different groups to observethe change of liver cells. Immunohistochemical results found that the positive stainingarea in AFB₁group is higher compared with the others groups. Cells were treated withaflatoxin B₁in different concentrations. The protein level of IDH3α was increasefollow the addition of aflatoxin. We choosed the optimal concentration of toxinaccording to the levels changes of IDH3α. IDH3α overexpression vector pCDNA3.1(-)-idh3α and four interference vectors with pSilencer4.1was constructed.Transfected these vector to mammalian cells by lipofection, then validated by WesternBlot.