Objective: To study the normal expression and location of wt-p53 protein on human high-metastasis hepatocellular carcinoma (HCC) cell line.
Methods: According to the sequence of human wild-type p53, the mp53 cDNA in which stopped codon TGA was mutated to TGG, was amplified by PCR, ligated to EGFP gene, of pEGFP-N1. The recombinant plasmid was transfected into MHCC97, in which p53 gene was mutated. The expression of human p53 protein was observed by fluorescence microscopy.
Results: There was a p53 mutation on 249 codon in human high-metastasis hepatocellular carcinoma cell line MHCC97, but no mutation in low-metastasis HCC cells LD20. The two genes of wt-p53 and GFP were fused in codon frame by DNA sequencing. The fluorescence microscopy showed that fluorescence light was spread in all of MHCC97 cells transfected with pEGFP-N1, but fluorescence light was focused on the nucleus of MHCC97 cells transfected with pEGFP-53.
Conclusion: The p53 mutation on 249 codon may be related to the character of metastasis of HCC cells. The human wild-type p53-GFP fusion protein is expressed in MHCC97 cell line and located in cell nucleus, which is similar to the expression of wt-p53 protein.