在不同條件下反應產生亞硝酸梅納反應產物(NOx-MRPs),在分別以4種NOx-MRPs(GL1、TL1、TL8、TH8)為起始劑(Initiator,I),及促進劑(Promoter,P),或以TPA當促進劑或以TPA當促進劑或以B(a)P當起始劑,來處理鼠纖維母細胞(C3H10T1/2)進行轉形作用,故而建立起亞硝酸化梅納反應所誘導的轉形(transformed)C3H10T1/2細胞10株,為了進一步探討其導致形細胞的原因,所有這些轉形(transformed)C3H10T1/2細胞進一步抽其RNA,合成cDNA,再以PCR-SSCP偵測p53之exon4--exon11及DNA定序反應偵測其p53抑癌基因是否突變,並探討導致亞硝酸化梅納反應物誘導轉形細胞的作用機制。發現所有10株細胞株裡,其p53突變比率為50%。而p53基因的突變位置大多位於exon7及exon10的區域。而突變大多以G為主要攻擊目標,而且突變型態有很多比例的G→A(Transition)突變。所以,得一結論知道亞硝酸梅納反應產物(NOx-MRPx)及TPA會誘發p53突變,導致C3H10T1/2細胞株轉形。
The N-nitro and N-nitroso Maillard reaction products(NOx-MRPs)in different condititon reaction,C3H10T1/2 cells were treated with the four kinds NOx-MRPs(TL1,TL1,TL8,TH8)which those as Initiator(I) or Promotor(P) or wiht TPA as Promotoror B(a)P as Initiator(I).We have been established the transformed C3H10T1/2 cell Iines by the N-nitro and N-nitrso Maillard reaction products.Why these 10transformed cell Iines can be induced The PCR-SSCP analysis exon4-exon11 of p53 and sequencing was used for screening and identificationthe p53 point mutation in the transformed cell Iines induced by NOx-MRPs or B(a)P or TPA products. The Mutation frequency of p53 was in 10 transformed
cell Iines induced by NOx-MRPs or B(a)P or TPA products is 50%.
P53 gene mutation was distributed on the exon7 and exon10.
Almost all the mutation were G→A transiton and the guanosine is the major target base. So we had a conclusion that the N-nitro and N-nitroso Maillard reaction Products and TPA can induced p53 mutation in the C3H10T1/2 cell lines.