Sequence analysis of the proximal promoter region of the human alpha-fetoprotein gene in hepatocellular carcinoma.
We have examined whether or not mutations exist in the proximal promoter region of the human alpha-fetoprotein (AFP) gene in the hepatocellular carcinoma (HCC) tissue. Genomic DNA was extracted from four patients: one HCC tissue, one HCC and its corresponding non-cancerous (cirrhosis) tissues, one liver cirrhosis (LC) tissue without HCC and one matching HCC tissue and peripheral blood leukocytes. Serum concentrations of AFP in the patients ranged from less than 5 to 10,138 ng/ml. Nucleotide sequence was determined by direct sequencing using a single-stranded DNA template that was produced first through the polymerase chain reaction (PCR) amplification and then asymmetric PCR. In one HCC tissue taken from the patient with a high concentration of serum AFP, nucleotides different from published ones were detected at -120 and -113. These changes, however, probably reflect a DNA polymorphism, because peripheral blood leukocytes of the same patient had the same changes. Including this patient, no mutations in the region from -160 to -10 were detected in the HCC specimens we have examined. These results suggest that the extremely proximal promoter region of the AFP gene where glucocorticoid-responsive element and HNF-1 binding sites exist is not responsible for the re-expression of AFP in HCC.
Igarashi
K
K
Department of Internal Medicine, Niigata University School of Medicine, Japan.
Aoyagi
Y
Y
Ohkoshi
S
S
Yokota
T
T
Mori
S
S
Suda
T
T
Kamimura
T
T
Asakura
H
H
D000328:Adult; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D005784:Gene Amplification / Q000235:genetics; D006801:Humans; D007962:Leukocytes / Q000502:physiology; D008103:Liver Cirrhosis / Q000097:blood / Q000235:genetics; D008113:Liver Neoplasms / Q000097:blood / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D000509:alpha-Fetoproteins / Q000235:genetics*
D004273:DNA, Neoplasm; D000509:alpha-Fetoproteins