[The expression and localization of wild-type p53-GFP fused gene on human high-metastasis hepatocellular carcinoma cell line].
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.


CONCLUSIONS
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.
He
P
P
Liver Cancer Institute and Zhong Shan Hospital, Shanghai Medical University, Shanghai 200032, China.
Tang
Z
Z
Ye
S
S
D019677:Artificial Gene Fusion; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000556:secondary; D003062:Codon; D016158:Genes, p53; D049452:Green Fluorescent Proteins; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics*; D009154:Mutation; D014407:Tumor Cells, Cultured
D003062:Codon; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins